Key Product Details

Validated by

Knockout/Knockdown, Biological Validation

Species Reactivity

Validated:

Mouse

Cited:

Mouse, Bovine, Transgenic Mouse

Applications

Validated:

Immunohistochemistry, Western Blot, Blockade of Receptor-ligand Interaction, Flow Cytometry, CyTOF-ready

Cited:

Immunohistochemistry, Immunohistochemistry-Frozen, Western Blot, Neutralization, Flow Cytometry, Immunocytochemistry, Immunoprecipitation, ELISA Development (Detection), In vivo assay

Label

Unconjugated

Antibody Source

Polyclonal Goat IgG
View all uPAR Primary Antibodies »

Product Specifications

Immunogen

Mouse myeloma cell line NS0-derived recombinant mouse uPAR isoform 1
Leu24-Thr297
Accession # Q545X5

Specificity

Detects mouse uPAR in direct ELISAs and Western blots.

Clonality

Polyclonal

Host

Goat

Isotype

IgG

Endotoxin Level

<0.10 EU per 1 μg of the antibody by the LAL method.

Scientific Data Images for Mouse uPAR Antibody

uPAR antibody in Mouse Kidney by Immunohistochemistry (IHC-Fr).

uPAR in Mouse Kidney.

uPAR was detected in perfusion fixed frozen sections of mouse kidney using Goat Anti-Mouse uPAR Antigen Affinity-purified Polyclonal Antibody (Catalog # AF534) at 3 µg/mL overnight at 4 °C. Tissue was stained using the Anti-Goat HRP-DAB Cell & Tissue Staining Kit (brown; CTS008) and counterstained with hematoxylin (blue). Specific staining was localized to plasma membrane in epithelial cells. View our protocol for Chromogenic IHC Staining of Frozen Tissue Sections.

Detection of uPAR in Raw264.7 cells by Flow Cytometry.

Raw264.7 cells were stained with Goat Anti-Mouse uPAR Antigen Affinity-purified Polyclonal Antibody (Catalog # AF534, filled histogram) or isotype control antibody (Catalog # AB-108-C, open histogram), followed by Phycoerythrin-conjugated Anti-Goat IgG Secondary Antibody (Catalog # F0107). View our protocol for Staining Membrane-associated Proteins.

Detection of Mouse uPAR by Flow Cytometry

Detection of Mouse uPAR by Flow Cytometry

Expression of murine Plaur in AT84 cells.In vitro characterization of AT84 cells stably transfected with either empty vector (EV) or a vector containing cDNA encoding murine uPAR (Plaur). A: Western blot analysis of whole cell lysates using a polyclonal anti-murine uPAR antibody (AF534). A total of 7.5 ng of recombinant murine uPAR (rmuPAR) was loaded as a positive control. Re-probing for beta -actin was used as a loading control. B: Western blot analysis of cellular membrane fractions using a polyclonal anti-murine uPAR antibody (AF534). Total protein was measured per sample and 53.5 µg of protein was loaded per lane. A and B: Images were cropped, as no additional bands were detectable. C: FACS analysis of non-permeabilized cells using a polyclonal anti-murine uPAR antibody (AF534). Alexa Fluor 488 anti-goat secondary antibody (A11055) was used as the secondary antibody. The quantified mean Alexa 488 fluorescence signal per cell line is presented in the panel to the right. D and E: Relative Plaur mRNA (uPAR) (D) or Plau mRNA (uPA) (E) expression levels as analysed using RT-qPCR. All expression levels were normalized to the expression of the reference genes Trfc and beta -actin. Error bars represent the standard error of mean (+SEM) and N = 3. One-way ANOVA; *p<0.05. F: Plasminogen-gelatin (upper panel) and gelatin (lower panel) zymography analysis of conditioned medium of cells cultured for 24 hours in SFM. HMW-uPA and mPLM (mouse plasmin) were loaded as positive controls. The images were cropped to size. G: Relative Plasminogen mRNA (Plg) expression levels as analysed using RT-qPCR. Error bars represent standard error of mean (+SEM) and N = 3. Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/25157856), licensed under a CC-BY license. Not internally tested by R&D Systems.
Detection of Mouse uPAR by Western Blot

Detection of Mouse uPAR by Western Blot

Leiomyoma conditioned medium induced uPAR expression.Analysis of uPAR expression induced by the LCM or purified ECM proteins in cultured uPAR knock-down cells. All Western blots were performed on whole cell lysates, and uPAR was detected using the polyclonal anti-murine uPAR antibody (AF534). A: Cells were either cultured in LCM (LM) or serum free medium (SF) for 24 hours. Cells were harvested with sample buffer and re-probing for beta -actin was used as a loading control. B: uPAR1-NT cells were seeded on different ECM protein substrates, incubated for 24 hours and harvested using RIPA buffer. 7.5 µg of total protein was loaded per lane. Equal loading was verified by re-probing for beta -actin. The poloxamer pluronic was used as a no-adhesion control. Col I = Collagen I, Vn = Vitronectin, Fn = Fibronectin, Lm = Laminin, ECL = Entactin, Collagen, Laminin, and FBS = Foetal Bovine Serum. C: Cells cultured in LCM (LM) or serum free medium (SF) for 24 hours were harvested using sample buffer and deglycosylated by PNGase F treatment (+) as indicated. Re-probing for beta -actin was used as a loading control. The three bands detected by the anti-uPAR antibody are labelled 1, 2, and 3, respectively. Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/25157856), licensed under a CC-BY license. Not internally tested by R&D Systems.
Detection of Mouse uPAR by Immunohistochemistry

Detection of Mouse uPAR by Immunohistochemistry

Tumour microenvironment induced uPAR protein expression in tongue tumours.Tumour growth pattern and uPAR protein levels in tongue tumours generated from the EV1, EV2, uPAR1 and uPAR2 cells. A–B: Representative images depicting the tumour growth pattern at the tumour-stroma interface in hematoxylin/eosin stained EV1 (A) and uPAR1 (B) tumours. Images were recorded at 10x magnification. C–D: Representative images depicting the IHC uPAR staining of the EV1 (C) or uPAR1 tumours (D). Images were recorded at 4x magnification. E–H: The images show high power magnification (20x magnifications) of the EV1 (E), uPAR1 (F), EV2 (G) and uPAR2 (H) tumours IHC stained for uPAR protein. Positive uPAR staining is seen as brown colour, and counterstaining was done with haematoxylin. I: The average staining index (SI) of the uPAR staining in the tumours. Maximum obtainable score is 9. The error bars shows the +SEM. N = number of tumours; EV1, N = 8/10; EV2, N = 5/10; uPAR1, N = 4/10; uPAR2 N = 9/10. One-way ANOVA; **p<0.01, *p<0.05. T = Tumours, S = Stroma. Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/25157856), licensed under a CC-BY license. Not internally tested by R&D Systems.
Detection of Mouse uPAR by Immunohistochemistry

Detection of Mouse uPAR by Immunohistochemistry

Leiomyoma stroma is a strong inducer of uPAR expression.Representative images of low- (EV1-sh3) and high- (uPAR1-NT) uPAR-expressing cells invading the ex vivo leiomyoma tissue. Cells were incubated for 7 and 14 days, as indicated. The tissue was IHC stained for uPAR. Positive uPAR staining is seen as brown colour, counterstained with haematoxylin. Images were recorded at 10x magnification. Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/25157856), licensed under a CC-BY license. Not internally tested by R&D Systems.
Detection of Mouse uPAR by Immunohistochemistry

Detection of Mouse uPAR by Immunohistochemistry

Tumour microenvironment induced uPAR protein expression in tongue tumours.Tumour growth pattern and uPAR protein levels in tongue tumours generated from the EV1, EV2, uPAR1 and uPAR2 cells. A–B: Representative images depicting the tumour growth pattern at the tumour-stroma interface in hematoxylin/eosin stained EV1 (A) and uPAR1 (B) tumours. Images were recorded at 10x magnification. C–D: Representative images depicting the IHC uPAR staining of the EV1 (C) or uPAR1 tumours (D). Images were recorded at 4x magnification. E–H: The images show high power magnification (20x magnifications) of the EV1 (E), uPAR1 (F), EV2 (G) and uPAR2 (H) tumours IHC stained for uPAR protein. Positive uPAR staining is seen as brown colour, and counterstaining was done with haematoxylin. I: The average staining index (SI) of the uPAR staining in the tumours. Maximum obtainable score is 9. The error bars shows the +SEM. N = number of tumours; EV1, N = 8/10; EV2, N = 5/10; uPAR1, N = 4/10; uPAR2 N = 9/10. One-way ANOVA; **p<0.01, *p<0.05. T = Tumours, S = Stroma. Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/25157856), licensed under a CC-BY license. Not internally tested by R&D Systems.
Detection of Mouse uPAR by Immunohistochemistry

Detection of Mouse uPAR by Immunohistochemistry

Tumour microenvironment induced uPAR protein expression in tongue tumours.Tumour growth pattern and uPAR protein levels in tongue tumours generated from the EV1, EV2, uPAR1 and uPAR2 cells. A–B: Representative images depicting the tumour growth pattern at the tumour-stroma interface in hematoxylin/eosin stained EV1 (A) and uPAR1 (B) tumours. Images were recorded at 10x magnification. C–D: Representative images depicting the IHC uPAR staining of the EV1 (C) or uPAR1 tumours (D). Images were recorded at 4x magnification. E–H: The images show high power magnification (20x magnifications) of the EV1 (E), uPAR1 (F), EV2 (G) and uPAR2 (H) tumours IHC stained for uPAR protein. Positive uPAR staining is seen as brown colour, and counterstaining was done with haematoxylin. I: The average staining index (SI) of the uPAR staining in the tumours. Maximum obtainable score is 9. The error bars shows the +SEM. N = number of tumours; EV1, N = 8/10; EV2, N = 5/10; uPAR1, N = 4/10; uPAR2 N = 9/10. One-way ANOVA; **p<0.01, *p<0.05. T = Tumours, S = Stroma. Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/25157856), licensed under a CC-BY license. Not internally tested by R&D Systems.
Detection of Mouse uPAR by Immunohistochemistry

Detection of Mouse uPAR by Immunohistochemistry

Tumour microenvironment induced uPAR protein expression in tongue tumours.Tumour growth pattern and uPAR protein levels in tongue tumours generated from the EV1, EV2, uPAR1 and uPAR2 cells. A–B: Representative images depicting the tumour growth pattern at the tumour-stroma interface in hematoxylin/eosin stained EV1 (A) and uPAR1 (B) tumours. Images were recorded at 10x magnification. C–D: Representative images depicting the IHC uPAR staining of the EV1 (C) or uPAR1 tumours (D). Images were recorded at 4x magnification. E–H: The images show high power magnification (20x magnifications) of the EV1 (E), uPAR1 (F), EV2 (G) and uPAR2 (H) tumours IHC stained for uPAR protein. Positive uPAR staining is seen as brown colour, and counterstaining was done with haematoxylin. I: The average staining index (SI) of the uPAR staining in the tumours. Maximum obtainable score is 9. The error bars shows the +SEM. N = number of tumours; EV1, N = 8/10; EV2, N = 5/10; uPAR1, N = 4/10; uPAR2 N = 9/10. One-way ANOVA; **p<0.01, *p<0.05. T = Tumours, S = Stroma. Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/25157856), licensed under a CC-BY license. Not internally tested by R&D Systems.
Detection of Mouse uPAR by Western Blot

Detection of Mouse uPAR by Western Blot

TGF-beta 1 reduces uPAR cleavage through induced PAI-1 expression. a, f-h. Western blot analysis of whole cell lysates of cultured and stimulated AT84-uPAR cells. Where indicated, cell lysates were either treated with PNGase F (+) or samples received the same treatment except that PNGase F was omitted (−). Glycosylated uPAR is indicated as uPARglc. d-e. Western blot analysis of conditioned medium from equally seeded amounts of AT84-EV and AT84-uPAR cells. a. Cells cultured in FBSM with or without 2 ng/ml active human TGF-beta 1 for 24 h. b-c. Total RNA from treated (TGF-beta 1) or untreated (Ctrl) cells was isolated and the relative expression of uPAR mRNA (B) or uPA mRNA (C) was analysed using RT-qPCR. The error bars show the +SD. N = 3. d. Cells cultured in SFM or FBSM and stimulated with 2 ng/ml TGF-beta 1 for 24 h as indicated and PAI-1 protein levels were analysed. Controls received either no additives (−) or the TGF-beta 1 buffer (0). Positive control: recombinant mouse PAI-1 (rmPAI-1). e. AT84-uPAR cells cultured in either SFM or in FBSM. Cells were either unstimulated (−) or stimulated (+) with 2 ng/ml TGF-beta 1 and/or 10 μM of the TGF-beta 1-inhibitor SB431542 as indicated. f. AT84-uPAR cells were cultured for 24 h in FBSM and stimulated with 2 ng/ml TGF-beta 1 as indicated. g. Deglycosylated whole cell lysates from AT84-uPAR cells cultured in SFM stimulated with 2 ng/ml TGF-beta 1 +/− the inhibitor SB431542 as indicated were analysed for uPAR protein levels. h. Cells were cultured for 24- or 48 h in SFM, FBSM or TMEM. Cells were treated with 2 ng/ml TGF-beta 1 in 10% FBS as indicated. The LRP1 protein is indicated. Arrowhead shows an unknown band. Image collected and cropped by CiteAb from the following open publication (https://bmccancer.biomedcentral.com/articles/10.1186/s12885-017-3349-7), licensed under a CC-BY license. Not internally tested by R&D Systems.
Detection of Mouse uPAR by Western Blot

Detection of Mouse uPAR by Western Blot

TGF-beta 1 reduces uPAR cleavage through induced PAI-1 expression. a, f-h. Western blot analysis of whole cell lysates of cultured and stimulated AT84-uPAR cells. Where indicated, cell lysates were either treated with PNGase F (+) or samples received the same treatment except that PNGase F was omitted (−). Glycosylated uPAR is indicated as uPARglc. d-e. Western blot analysis of conditioned medium from equally seeded amounts of AT84-EV and AT84-uPAR cells. a. Cells cultured in FBSM with or without 2 ng/ml active human TGF-beta 1 for 24 h. b-c. Total RNA from treated (TGF-beta 1) or untreated (Ctrl) cells was isolated and the relative expression of uPAR mRNA (B) or uPA mRNA (C) was analysed using RT-qPCR. The error bars show the +SD. N = 3. d. Cells cultured in SFM or FBSM and stimulated with 2 ng/ml TGF-beta 1 for 24 h as indicated and PAI-1 protein levels were analysed. Controls received either no additives (−) or the TGF-beta 1 buffer (0). Positive control: recombinant mouse PAI-1 (rmPAI-1). e. AT84-uPAR cells cultured in either SFM or in FBSM. Cells were either unstimulated (−) or stimulated (+) with 2 ng/ml TGF-beta 1 and/or 10 μM of the TGF-beta 1-inhibitor SB431542 as indicated. f. AT84-uPAR cells were cultured for 24 h in FBSM and stimulated with 2 ng/ml TGF-beta 1 as indicated. g. Deglycosylated whole cell lysates from AT84-uPAR cells cultured in SFM stimulated with 2 ng/ml TGF-beta 1 +/− the inhibitor SB431542 as indicated were analysed for uPAR protein levels. h. Cells were cultured for 24- or 48 h in SFM, FBSM or TMEM. Cells were treated with 2 ng/ml TGF-beta 1 in 10% FBS as indicated. The LRP1 protein is indicated. Arrowhead shows an unknown band. Image collected and cropped by CiteAb from the following open publication (https://bmccancer.biomedcentral.com/articles/10.1186/s12885-017-3349-7), licensed under a CC-BY license. Not internally tested by R&D Systems.
Detection of Mouse uPAR by Immunohistochemistry

Detection of Mouse uPAR by Immunohistochemistry

Tumour microenvironment induced uPAR protein expression in tongue tumours.Tumour growth pattern and uPAR protein levels in tongue tumours generated from the EV1, EV2, uPAR1 and uPAR2 cells. A–B: Representative images depicting the tumour growth pattern at the tumour-stroma interface in hematoxylin/eosin stained EV1 (A) and uPAR1 (B) tumours. Images were recorded at 10x magnification. C–D: Representative images depicting the IHC uPAR staining of the EV1 (C) or uPAR1 tumours (D). Images were recorded at 4x magnification. E–H: The images show high power magnification (20x magnifications) of the EV1 (E), uPAR1 (F), EV2 (G) and uPAR2 (H) tumours IHC stained for uPAR protein. Positive uPAR staining is seen as brown colour, and counterstaining was done with haematoxylin. I: The average staining index (SI) of the uPAR staining in the tumours. Maximum obtainable score is 9. The error bars shows the +SEM. N = number of tumours; EV1, N = 8/10; EV2, N = 5/10; uPAR1, N = 4/10; uPAR2 N = 9/10. One-way ANOVA; **p<0.01, *p<0.05. T = Tumours, S = Stroma. Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/25157856), licensed under a CC-BY license. Not internally tested by R&D Systems.
Detection of Mouse uPAR by Western Blot

Detection of Mouse uPAR by Western Blot

Leiomyoma conditioned medium induced uPAR expression.Analysis of uPAR expression induced by the LCM or purified ECM proteins in cultured uPAR knock-down cells. All Western blots were performed on whole cell lysates, and uPAR was detected using the polyclonal anti-murine uPAR antibody (AF534). A: Cells were either cultured in LCM (LM) or serum free medium (SF) for 24 hours. Cells were harvested with sample buffer and re-probing for beta -actin was used as a loading control. B: uPAR1-NT cells were seeded on different ECM protein substrates, incubated for 24 hours and harvested using RIPA buffer. 7.5 µg of total protein was loaded per lane. Equal loading was verified by re-probing for beta -actin. The poloxamer pluronic was used as a no-adhesion control. Col I = Collagen I, Vn = Vitronectin, Fn = Fibronectin, Lm = Laminin, ECL = Entactin, Collagen, Laminin, and FBS = Foetal Bovine Serum. C: Cells cultured in LCM (LM) or serum free medium (SF) for 24 hours were harvested using sample buffer and deglycosylated by PNGase F treatment (+) as indicated. Re-probing for beta -actin was used as a loading control. The three bands detected by the anti-uPAR antibody are labelled 1, 2, and 3, respectively. Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/25157856), licensed under a CC-BY license. Not internally tested by R&D Systems.
Detection of Mouse uPAR by Western Blot

Detection of Mouse uPAR by Western Blot

Expression of murine Plaur in AT84 cells.In vitro characterization of AT84 cells stably transfected with either empty vector (EV) or a vector containing cDNA encoding murine uPAR (Plaur). A: Western blot analysis of whole cell lysates using a polyclonal anti-murine uPAR antibody (AF534). A total of 7.5 ng of recombinant murine uPAR (rmuPAR) was loaded as a positive control. Re-probing for beta -actin was used as a loading control. B: Western blot analysis of cellular membrane fractions using a polyclonal anti-murine uPAR antibody (AF534). Total protein was measured per sample and 53.5 µg of protein was loaded per lane. A and B: Images were cropped, as no additional bands were detectable. C: FACS analysis of non-permeabilized cells using a polyclonal anti-murine uPAR antibody (AF534). Alexa Fluor 488 anti-goat secondary antibody (A11055) was used as the secondary antibody. The quantified mean Alexa 488 fluorescence signal per cell line is presented in the panel to the right. D and E: Relative Plaur mRNA (uPAR) (D) or Plau mRNA (uPA) (E) expression levels as analysed using RT-qPCR. All expression levels were normalized to the expression of the reference genes Trfc and beta -actin. Error bars represent the standard error of mean (+SEM) and N = 3. One-way ANOVA; *p<0.05. F: Plasminogen-gelatin (upper panel) and gelatin (lower panel) zymography analysis of conditioned medium of cells cultured for 24 hours in SFM. HMW-uPA and mPLM (mouse plasmin) were loaded as positive controls. The images were cropped to size. G: Relative Plasminogen mRNA (Plg) expression levels as analysed using RT-qPCR. Error bars represent standard error of mean (+SEM) and N = 3. Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/25157856), licensed under a CC-BY license. Not internally tested by R&D Systems.
Detection of Mouse uPAR by Immunohistochemistry

Detection of Mouse uPAR by Immunohistochemistry

Tumour microenvironment induced uPAR protein expression in tongue tumours.Tumour growth pattern and uPAR protein levels in tongue tumours generated from the EV1, EV2, uPAR1 and uPAR2 cells. A–B: Representative images depicting the tumour growth pattern at the tumour-stroma interface in hematoxylin/eosin stained EV1 (A) and uPAR1 (B) tumours. Images were recorded at 10x magnification. C–D: Representative images depicting the IHC uPAR staining of the EV1 (C) or uPAR1 tumours (D). Images were recorded at 4x magnification. E–H: The images show high power magnification (20x magnifications) of the EV1 (E), uPAR1 (F), EV2 (G) and uPAR2 (H) tumours IHC stained for uPAR protein. Positive uPAR staining is seen as brown colour, and counterstaining was done with haematoxylin. I: The average staining index (SI) of the uPAR staining in the tumours. Maximum obtainable score is 9. The error bars shows the +SEM. N = number of tumours; EV1, N = 8/10; EV2, N = 5/10; uPAR1, N = 4/10; uPAR2 N = 9/10. One-way ANOVA; **p<0.01, *p<0.05. T = Tumours, S = Stroma. Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/25157856), licensed under a CC-BY license. Not internally tested by R&D Systems.
Detection of Mouse uPAR by Western Blot

Detection of Mouse uPAR by Western Blot

Soluble uPAR shed from mouse OSCC cells induce migration. a-b. Western blot analysis of uPAR protein levels in concentrated conditioned medium from AT84-EV and AT84-uPAR cells. a. Conditioned medium from cells cultured in SFM or SFM containing 10 nM plasminogen (plg) as indicated. The conditioned medium was concentrated from 2 ml to 110 μl for both samples, and deglycosylated using PNGase F. b. Conditioned medium from cells cultured in SFM or in FBSM was concentrated from 2 ml to 350 μl for each sample. A whole cell lysate sample of AT84-uPAR cells was used as a positive control. The two indicated FBSM samples are replicates. Glycosylated suPAR is indicated as suPARglc. c. Non-concentrated conditioned medium from AT84-EV (EV medium) and AT84-uPAR cells (uPAR-medium) was used as an attractant for the AT84-EV cells in a real-time cell migration analysis using the xCELLigence system. N = 3, where each experiment had two technical replicates, and the error bars represent the standard error of mean (±SEM) based on three separate experiments. Image collected and cropped by CiteAb from the following open publication (https://bmccancer.biomedcentral.com/articles/10.1186/s12885-017-3349-7), licensed under a CC-BY license. Not internally tested by R&D Systems.
Detection of Mouse uPAR by Immunohistochemistry

Detection of Mouse uPAR by Immunohistochemistry

In vivo tongue tumours of EV1 and uPAR1 knock-down cells.IHC uPAR staining and growth pattern of tongue tumours generated from the EV1 and uPAR1 cells containing either shRNA targeting uPAR (EV1-sh and uPAR1-sh), or non-targeting shRNA (uPAR1-NT). A–C: IHC uPAR staining of EV1-sh (A), uPAR1-NT (B) and uPAR1-sh (C) tumours, respectively. Images were recorded at 20x magnifications. D: Representative image depicting the tumour growth pattern at the tumour-stroma interface in hematoxylin/eosin stained EV1-sh. E: The average SI of the uPAR staining in the tumours, with the maximum obtainable score of 9. The error bars shows the +SEM. N = number of tumours; EV1, N = 8/10; EV1-sh, N = 11/16; uPAR1, N = 4/10; uPAR1-NT, N = 3/8; uPAR1-sh, N = 4/16. One-way ANOVA; **p<0.01, *p<0.05. T = Tumour, S = Stroma. Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/25157856), licensed under a CC-BY license. Not internally tested by R&D Systems.
Detection of Mouse uPAR by Immunohistochemistry

Detection of Mouse uPAR by Immunohistochemistry

In vivo tongue tumours of EV1 and uPAR1 knock-down cells.IHC uPAR staining and growth pattern of tongue tumours generated from the EV1 and uPAR1 cells containing either shRNA targeting uPAR (EV1-sh and uPAR1-sh), or non-targeting shRNA (uPAR1-NT). A–C: IHC uPAR staining of EV1-sh (A), uPAR1-NT (B) and uPAR1-sh (C) tumours, respectively. Images were recorded at 20x magnifications. D: Representative image depicting the tumour growth pattern at the tumour-stroma interface in hematoxylin/eosin stained EV1-sh. E: The average SI of the uPAR staining in the tumours, with the maximum obtainable score of 9. The error bars shows the +SEM. N = number of tumours; EV1, N = 8/10; EV1-sh, N = 11/16; uPAR1, N = 4/10; uPAR1-NT, N = 3/8; uPAR1-sh, N = 4/16. One-way ANOVA; **p<0.01, *p<0.05. T = Tumour, S = Stroma. Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/25157856), licensed under a CC-BY license. Not internally tested by R&D Systems.
Detection of Mouse uPAR by Western Blot

Detection of Mouse uPAR by Western Blot

Knock-down of uPAR expression in AT84 cells.shRNA knock-down of Plaur in AT84 cells. A: Flow chart showing the generation of the single cell clones. B: Western blot analysis of whole cell lysates from the single cell clones stably transfected with either shRNA-constructs (shRNA 3 = sh3, shRNA 4 = sh4 or shRNA 5 = sh5) targeting Plaur or constructs containing non-target shRNA (NT) or the empty vector (EV). uPAR was detected using a polyclonal anti-murine uPAR antibody (AF534). Re-probing for beta -actin was used as a loading control. Images were cropped, as no additional bands were detected in the blot. Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/25157856), licensed under a CC-BY license. Not internally tested by R&D Systems.
Detection of Mouse uPAR by Western Blot

Detection of Mouse uPAR by Western Blot

TGF-beta 1 reduces uPAR cleavage through induced PAI-1 expression. a, f-h. Western blot analysis of whole cell lysates of cultured and stimulated AT84-uPAR cells. Where indicated, cell lysates were either treated with PNGase F (+) or samples received the same treatment except that PNGase F was omitted (−). Glycosylated uPAR is indicated as uPARglc. d-e. Western blot analysis of conditioned medium from equally seeded amounts of AT84-EV and AT84-uPAR cells. a. Cells cultured in FBSM with or without 2 ng/ml active human TGF-beta 1 for 24 h. b-c. Total RNA from treated (TGF-beta 1) or untreated (Ctrl) cells was isolated and the relative expression of uPAR mRNA (B) or uPA mRNA (C) was analysed using RT-qPCR. The error bars show the +SD. N = 3. d. Cells cultured in SFM or FBSM and stimulated with 2 ng/ml TGF-beta 1 for 24 h as indicated and PAI-1 protein levels were analysed. Controls received either no additives (−) or the TGF-beta 1 buffer (0). Positive control: recombinant mouse PAI-1 (rmPAI-1). e. AT84-uPAR cells cultured in either SFM or in FBSM. Cells were either unstimulated (−) or stimulated (+) with 2 ng/ml TGF-beta 1 and/or 10 μM of the TGF-beta 1-inhibitor SB431542 as indicated. f. AT84-uPAR cells were cultured for 24 h in FBSM and stimulated with 2 ng/ml TGF-beta 1 as indicated. g. Deglycosylated whole cell lysates from AT84-uPAR cells cultured in SFM stimulated with 2 ng/ml TGF-beta 1 +/− the inhibitor SB431542 as indicated were analysed for uPAR protein levels. h. Cells were cultured for 24- or 48 h in SFM, FBSM or TMEM. Cells were treated with 2 ng/ml TGF-beta 1 in 10% FBS as indicated. The LRP1 protein is indicated. Arrowhead shows an unknown band. Image collected and cropped by CiteAb from the following open publication (https://bmccancer.biomedcentral.com/articles/10.1186/s12885-017-3349-7), licensed under a CC-BY license. Not internally tested by R&D Systems.
Detection of Mouse uPAR by Western Blot

Detection of Mouse uPAR by Western Blot

Soluble uPAR shed from mouse OSCC cells induce migration. a-b. Western blot analysis of uPAR protein levels in concentrated conditioned medium from AT84-EV and AT84-uPAR cells. a. Conditioned medium from cells cultured in SFM or SFM containing 10 nM plasminogen (plg) as indicated. The conditioned medium was concentrated from 2 ml to 110 μl for both samples, and deglycosylated using PNGase F. b. Conditioned medium from cells cultured in SFM or in FBSM was concentrated from 2 ml to 350 μl for each sample. A whole cell lysate sample of AT84-uPAR cells was used as a positive control. The two indicated FBSM samples are replicates. Glycosylated suPAR is indicated as suPARglc. c. Non-concentrated conditioned medium from AT84-EV (EV medium) and AT84-uPAR cells (uPAR-medium) was used as an attractant for the AT84-EV cells in a real-time cell migration analysis using the xCELLigence system. N = 3, where each experiment had two technical replicates, and the error bars represent the standard error of mean (±SEM) based on three separate experiments. Image collected and cropped by CiteAb from the following open publication (https://bmccancer.biomedcentral.com/articles/10.1186/s12885-017-3349-7), licensed under a CC-BY license. Not internally tested by R&D Systems.
Detection of Mouse uPAR by Western Blot

Detection of Mouse uPAR by Western Blot

Expression of murine Plaur in AT84 cells.In vitro characterization of AT84 cells stably transfected with either empty vector (EV) or a vector containing cDNA encoding murine uPAR (Plaur). A: Western blot analysis of whole cell lysates using a polyclonal anti-murine uPAR antibody (AF534). A total of 7.5 ng of recombinant murine uPAR (rmuPAR) was loaded as a positive control. Re-probing for beta -actin was used as a loading control. B: Western blot analysis of cellular membrane fractions using a polyclonal anti-murine uPAR antibody (AF534). Total protein was measured per sample and 53.5 µg of protein was loaded per lane. A and B: Images were cropped, as no additional bands were detectable. C: FACS analysis of non-permeabilized cells using a polyclonal anti-murine uPAR antibody (AF534). Alexa Fluor 488 anti-goat secondary antibody (A11055) was used as the secondary antibody. The quantified mean Alexa 488 fluorescence signal per cell line is presented in the panel to the right. D and E: Relative Plaur mRNA (uPAR) (D) or Plau mRNA (uPA) (E) expression levels as analysed using RT-qPCR. All expression levels were normalized to the expression of the reference genes Trfc and beta -actin. Error bars represent the standard error of mean (+SEM) and N = 3. One-way ANOVA; *p<0.05. F: Plasminogen-gelatin (upper panel) and gelatin (lower panel) zymography analysis of conditioned medium of cells cultured for 24 hours in SFM. HMW-uPA and mPLM (mouse plasmin) were loaded as positive controls. The images were cropped to size. G: Relative Plasminogen mRNA (Plg) expression levels as analysed using RT-qPCR. Error bars represent standard error of mean (+SEM) and N = 3. Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/25157856), licensed under a CC-BY license. Not internally tested by R&D Systems.
Detection of Mouse uPAR by Western Blot

Detection of Mouse uPAR by Western Blot

Leiomyoma conditioned medium induced uPAR expression.Analysis of uPAR expression induced by the LCM or purified ECM proteins in cultured uPAR knock-down cells. All Western blots were performed on whole cell lysates, and uPAR was detected using the polyclonal anti-murine uPAR antibody (AF534). A: Cells were either cultured in LCM (LM) or serum free medium (SF) for 24 hours. Cells were harvested with sample buffer and re-probing for beta -actin was used as a loading control. B: uPAR1-NT cells were seeded on different ECM protein substrates, incubated for 24 hours and harvested using RIPA buffer. 7.5 µg of total protein was loaded per lane. Equal loading was verified by re-probing for beta -actin. The poloxamer pluronic was used as a no-adhesion control. Col I = Collagen I, Vn = Vitronectin, Fn = Fibronectin, Lm = Laminin, ECL = Entactin, Collagen, Laminin, and FBS = Foetal Bovine Serum. C: Cells cultured in LCM (LM) or serum free medium (SF) for 24 hours were harvested using sample buffer and deglycosylated by PNGase F treatment (+) as indicated. Re-probing for beta -actin was used as a loading control. The three bands detected by the anti-uPAR antibody are labelled 1, 2, and 3, respectively. Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/25157856), licensed under a CC-BY license. Not internally tested by R&D Systems.
Detection of Mouse uPAR by Immunohistochemistry

Detection of Mouse uPAR by Immunohistochemistry

In vivo tongue tumours of EV1 and uPAR1 knock-down cells.IHC uPAR staining and growth pattern of tongue tumours generated from the EV1 and uPAR1 cells containing either shRNA targeting uPAR (EV1-sh and uPAR1-sh), or non-targeting shRNA (uPAR1-NT). A–C: IHC uPAR staining of EV1-sh (A), uPAR1-NT (B) and uPAR1-sh (C) tumours, respectively. Images were recorded at 20x magnifications. D: Representative image depicting the tumour growth pattern at the tumour-stroma interface in hematoxylin/eosin stained EV1-sh. E: The average SI of the uPAR staining in the tumours, with the maximum obtainable score of 9. The error bars shows the +SEM. N = number of tumours; EV1, N = 8/10; EV1-sh, N = 11/16; uPAR1, N = 4/10; uPAR1-NT, N = 3/8; uPAR1-sh, N = 4/16. One-way ANOVA; **p<0.01, *p<0.05. T = Tumour, S = Stroma. Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/25157856), licensed under a CC-BY license. Not internally tested by R&D Systems.
Detection of Mouse uPAR by Western Blot

Detection of Mouse uPAR by Western Blot

Expression of murine Plaur in AT84 cells.In vitro characterization of AT84 cells stably transfected with either empty vector (EV) or a vector containing cDNA encoding murine uPAR (Plaur). A: Western blot analysis of whole cell lysates using a polyclonal anti-murine uPAR antibody (AF534). A total of 7.5 ng of recombinant murine uPAR (rmuPAR) was loaded as a positive control. Re-probing for beta -actin was used as a loading control. B: Western blot analysis of cellular membrane fractions using a polyclonal anti-murine uPAR antibody (AF534). Total protein was measured per sample and 53.5 µg of protein was loaded per lane. A and B: Images were cropped, as no additional bands were detectable. C: FACS analysis of non-permeabilized cells using a polyclonal anti-murine uPAR antibody (AF534). Alexa Fluor 488 anti-goat secondary antibody (A11055) was used as the secondary antibody. The quantified mean Alexa 488 fluorescence signal per cell line is presented in the panel to the right. D and E: Relative Plaur mRNA (uPAR) (D) or Plau mRNA (uPA) (E) expression levels as analysed using RT-qPCR. All expression levels were normalized to the expression of the reference genes Trfc and beta -actin. Error bars represent the standard error of mean (+SEM) and N = 3. One-way ANOVA; *p<0.05. F: Plasminogen-gelatin (upper panel) and gelatin (lower panel) zymography analysis of conditioned medium of cells cultured for 24 hours in SFM. HMW-uPA and mPLM (mouse plasmin) were loaded as positive controls. The images were cropped to size. G: Relative Plasminogen mRNA (Plg) expression levels as analysed using RT-qPCR. Error bars represent standard error of mean (+SEM) and N = 3. Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/25157856), licensed under a CC-BY license. Not internally tested by R&D Systems.
Detection of Mouse Mouse uPAR Antibody by Western Blot

Detection of Mouse Mouse uPAR Antibody by Western Blot

Soluble uPAR shed from mouse OSCC cells induce migration. a-b. Western blot analysis of uPAR protein levels in concentrated conditioned medium from AT84-EV and AT84-uPAR cells. a. Conditioned medium from cells cultured in SFM or SFM containing 10 nM plasminogen (plg) as indicated. The conditioned medium was concentrated from 2 ml to 110 μl for both samples, and deglycosylated using PNGase F. b. Conditioned medium from cells cultured in SFM or in FBSM was concentrated from 2 ml to 350 μl for each sample. A whole cell lysate sample of AT84-uPAR cells was used as a positive control. The two indicated FBSM samples are replicates. Glycosylated suPAR is indicated as suPARglc. c. Non-concentrated conditioned medium from AT84-EV (EV medium) and AT84-uPAR cells (uPAR-medium) was used as an attractant for the AT84-EV cells in a real-time cell migration analysis using the xCELLigence system. N = 3, where each experiment had two technical replicates, and the error bars represent the standard error of mean (±SEM) based on three separate experiments Image collected and cropped by CiteAb from the following publication (https://pubmed.ncbi.nlm.nih.gov/28526008), licensed under a CC-BY license. Not internally tested by R&D Systems.

Applications for Mouse uPAR Antibody

Application
Recommended Usage

Blockade of Receptor-ligand Interaction

In a functional ELISA, 0.8-4 µg/mL of this antibody will block 50% of the binding of 50 ng/mL of Recombinant Human u-Plasminogen Activator/Urokinase (Catalog # 1310-SE) to immobilized Recombinant Mouse uPAR Fc Chimera (Catalog # 531-PA) coated at 5 µg/mL (100 µL/well). At 25 μg/mL, this antibody will block >90% of the binding.

CyTOF-ready

Ready to be labeled using established conjugation methods. No BSA or other carrier proteins that could interfere with conjugation.

Flow Cytometry

0.25 µg/106 cells
Sample: RAW 264.7 mouse monocyte/macrophage cell line

Immunohistochemistry

5-15 µg/mL
Sample: Perfusion fixed frozen sections of mouse kidney

Western Blot

0.1 µg/mL
Sample: Recombinant Mouse uPAR Fc Chimera (Catalog # 531-PA)

Reviewed Applications

Read 1 review rated 5 using AF534 in the following applications:

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Formulation, Preparation, and Storage

Purification

Antigen Affinity-purified

Reconstitution

Reconstitute at 0.2 mg/mL in sterile PBS. For liquid material, refer to CoA for concentration.

Formulation

Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. *Small pack size (SP) is supplied either lyophilized or as a 0.2 µm filtered solution in PBS.

Shipping

Lyophilized product is shipped at ambient temperature. Liquid small pack size (-SP) is shipped with polar packs. Upon receipt, store immediately at the temperature recommended below.

Stability & Storage

Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
  • 12 months from date of receipt, -20 to -70 °C as supplied.
  • 1 month, 2 to 8 °C under sterile conditions after reconstitution.
  • 6 months, -20 to -70 °C under sterile conditions after reconstitution.

Calculators

The reconstitution calculator allows you to quickly calculate the volume of a reagent to reconstitute your vial. Simply enter the mass of reagent and the target concentration and the calculator will determine the rest.

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Background: uPAR

The urokinase-type plasminogen activator (uPA) is one of two activators that converts the extracellular zymogen plasminogen to plasmin, a serine protease that is involved in a variety of normal and pathological processes that require cell migration and/or tissue destruction. uPA is synthesized and released from cells as a single‑chain (sc) pro-enzyme with limited enzymatic activity and is converted to an active two-chain (tc) disulfide-linked active enzyme by plasmin and other specific proteinases. Both the scuPA and tcuPA bind with high-affinity to the cell surface via the glycosyl phosphatidylinositol-linked receptor uPAR which serves to localize the uPA proteolytic activity. The enzymatic activity of scuPA has also been shown to be enhanced by binding to uPAR. Independent of their proteolytic activity, the uPA/uPAR interaction also initiates signal transduction responses resulting in activation of protein tyrosine kinases, gene expression, cell adhesion, and chemotaxis. uPAR can interact with integrins to suppress normal integrin adhesive function and promote adhesion to vitronectin through a high affinity vitronectin binding site on uPAR. Mouse uPAR-1/Fc cDNA encodes a 327 amino acid (aa) residue precursor protein with a 23 aa residue signal peptide, seven potential N-linked glycosylation sites and a C-terminal GPI-anchor site. An alternate spliced variant of uPAR encoding a secreted soluble form of uPAR also exists. Human and mouse uPAR share approximately 60% aa sequence identity and the receptor-ligand interaction is highly species-specific. Human uPA binds rmuPAR at a lower affinity compared to rhuPAR.

References

  1. Dear, A.E. and R.L. Medcalf (1998) Eur. J. Biochemistry 252:185.

Long Name

Urokinase-type Plasminogen Activator Receptor

Alternate Names

PLAUR

Entrez Gene IDs

5329 (Human); 18793 (Mouse); 102139334 (Cynomolgus Monkey)

Gene Symbol

PLAUR

UniProt

Q545X5

Additional uPAR Products

Product Documents for Mouse uPAR Antibody

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Product Specific Notices for Mouse uPAR Antibody

For research use only

Citations for Mouse uPAR Antibody

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  • Mouse uPAR Antibody
    Name: Anonymous
    Application: Immunocytochemistry/Immunofluorescence
    Sample Tested: bEnd.3 mouse endothelioma cell line
    Species: Mouse
    Verified Customer | Posted 01/13/2022
    No permeabilization with triton X-100 1% BSA, 30 min, RT Primary Ab: 1 ug/mL, 1h, RT Secondary Ab: AlexaFluor 448 donkey anti-goat IgG (H+L), 1:800, 30 min, RT
    Mouse uPAR Antibody AF534

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