Human Serpin E1/PAI-1 Antibody

R&D Systems | Catalog # AF1786

R&D Systems

Key Product Details

Species Reactivity

Validated:

Human

Cited:

Human, Mouse, Rat, Xenograft

Applications

Validated:

Immunohistochemistry, Western Blot, Neutralization, Simple Western, Immunoprecipitation

Cited:

Immunohistochemistry, Immunohistochemistry-Frozen, Western Blot, Neutralization, Dot Blot

Label

Unconjugated

Antibody Source

Polyclonal Goat IgG
View all Serpin E1/PAI-1 Primary Antibodies »

Product Specifications

Immunogen

S. frugiperda insect ovarian cell line Sf 21-derived recombinant human Serpin E1/PAI‑1
Gly21-Pro402
Accession # P05121

Specificity

Detects human Serpin E1/PAI‑1 in direct ELISAs and Western blots. In direct ELISAs, approximately 15% cross-reactivity with recombinant mouse Serpin E1 is observed.

Clonality

Polyclonal

Host

Goat

Isotype

IgG

Scientific Data Images for Human Serpin E1/PAI-1 Antibody

Detection of Human Serpin E1/PAI-1 antibody by Western Blot.

Detection of Human Serpin E1/PAI‑1 by Western Blot.

Western blot shows lysates of HUVEC human umbilical vein endothelial cells. PVDF membrane was probed with 1 µg/mL of Goat Anti-Human Serpin E1/PAI-1 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF1786) followed by HRP-conjugated Anti-Goat IgG Secondary Antibody (Catalog # HAF017). A specific band was detected for Serpin E1/PAI-1 at approximately 45 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.
Serpin E1/PAI-1 antibody in Human Liver Cancer Tissue by Immunohistochemistry (IHC-P).

Serpin E1/PAI‑1 in Human Liver Cancer Tissue.

Serpin E1/PAI-1 was detected in immersion fixed paraffin-embedded sections of human liver cancer tissue using Goat Anti-Human Serpin E1/PAI-1 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF1786) at 15 µg/mL overnight at 4 °C. Tissue was stained using the Anti-Goat HRP-DAB Cell & Tissue Staining Kit (brown; Catalog # CTS008) and counterstained with hematoxylin (blue). View our protocol for Chromogenic IHC Staining of Paraffin-embedded Tissue Sections.
Detection of Human Serpin E1/PAI-1 antibody by Simple WesternTM.

Detection of Human Serpin E1/PAI‑1 by Simple WesternTM.

Simple Western lane view shows lysates of HUVEC human umbilical vein endothelial cells, loaded at 0.2 mg/mL. A specific band was detected for Serpin E1/PAI‑1 at approximately 54 kDa (as indicated) using 2.5 µg/mL of Goat Anti-Human Serpin E1/PAI‑1 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF1786) followed by 1:50 dilution of HRP-conjugated Anti-Goat IgG Secondary Antibody (Catalog # HAF109). This experiment was conducted under reducing conditions and using the 12-230 kDa separation system.
Neutralization of Serpin E1/ PAI‑1 Activity by Human Serpin E1/PAI‑1 Antibody.

Neutralization of Serpin E1/ PAI‑1 Activity by Human Serpin E1/PAI‑1 Antibody.

Recombinant Human u-Plasminogen Activator (uPA)/Urokinase (0.1 µg/mL, Catalog # 1310-SE) activity is measured in the presence of Recombinant Human Serpin E1 (1.35 µg/mL, Catalog # 1786-PI) that has been preincubated with increasing concentrations of Goat Anti-Human Serpin E1/PAI-1 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF1786). The ND50 is typically 10 µg/mL.
Detection of Serpin E1/PAI-1 by Immunohistochemistry

Detection of Serpin E1/PAI-1 by Immunohistochemistry

The expression of PAI-1 and LRP1 in human ESCC tissues. A Immunohistochemical staining for PAI-1 in human ESCC tissues. Representative images are shown: PAI-1 low-intensity in the stroma (left) and PAI-1 high-intensity in the stroma (right). Scale bars = 100 μm. b High-power fields of immunohistochemical staining for PAI-1 in same cases as Fig. 6a. Scale bars = 20 μm. Arrow heads: vascular endothelial cell. c Immunohistochemical staining for LRP1 in cancer nest of human ESCC tissues. Representative images are shown: LRP1 low-intensity in cancer nest (left) and LRP1 high-intensity in cancer nest (right). Scale bars = 100 μm. d Immunohistochemical staining for LRP1 in the stroma of human ESCC tissues. Representative images are shown: LRP1 low-intensity in the stroma (left) and LRP1 high-intensity in the stroma (right). Scale bars = 20 μm. e Stromal cells, including CD204+ TAMs, expressed LRP1 in human ESCC tissue. Double immunofluorescence was performed using anti-LRP1 (green) and anti-CD204 (red) antibodies in formalin-fixed paraffin-embedded section of human ESCC tissue. LRP1 was expressed in both cancer cells and stromal cells, and some of the LRP1-positive stromal cells were CD204+ macrophages (arrow heads). Nuclei were stained with DAPI (blue). Scale bar = 20 μm. CA cancer nest. ST stroma. Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/33311557), licensed under a CC-BY license. Not internally tested by R&D Systems.
Detection of Serpin E1/PAI-1 by Immunohistochemistry

Detection of Serpin E1/PAI-1 by Immunohistochemistry

The expression of PAI-1 and LRP1 in human ESCC tissues. A Immunohistochemical staining for PAI-1 in human ESCC tissues. Representative images are shown: PAI-1 low-intensity in the stroma (left) and PAI-1 high-intensity in the stroma (right). Scale bars = 100 μm. b High-power fields of immunohistochemical staining for PAI-1 in same cases as Fig. 6a. Scale bars = 20 μm. Arrow heads: vascular endothelial cell. c Immunohistochemical staining for LRP1 in cancer nest of human ESCC tissues. Representative images are shown: LRP1 low-intensity in cancer nest (left) and LRP1 high-intensity in cancer nest (right). Scale bars = 100 μm. d Immunohistochemical staining for LRP1 in the stroma of human ESCC tissues. Representative images are shown: LRP1 low-intensity in the stroma (left) and LRP1 high-intensity in the stroma (right). Scale bars = 20 μm. e Stromal cells, including CD204+ TAMs, expressed LRP1 in human ESCC tissue. Double immunofluorescence was performed using anti-LRP1 (green) and anti-CD204 (red) antibodies in formalin-fixed paraffin-embedded section of human ESCC tissue. LRP1 was expressed in both cancer cells and stromal cells, and some of the LRP1-positive stromal cells were CD204+ macrophages (arrow heads). Nuclei were stained with DAPI (blue). Scale bar = 20 μm. CA cancer nest. ST stroma. Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/33311557), licensed under a CC-BY license. Not internally tested by R&D Systems.
Human Serpin E1 / PAI-1 ELISA Standard Curve

Human Serpin E1 / PAI-1 ELISA Standard Curve

Recombinant Human Serpin E1/PAI‑1 (Catalog # 1786-PI) was serially diluted and captured by Mouse Anti-Human Serpin E1/PAI‑1 Monoclonal Antibody (Catalog # MAB1786) coated on a Clear Polystyrene Microplate (Catalog # DY990). Goat Anti-Human Serpin E1/PAI‑1 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF1786) was biotinylated and incubated with the protein captured on the plate. Detection of the standard curve was achieved by incubating Streptavidin-HRP (Catalog # DY998)

Applications for Human Serpin E1/PAI-1 Antibody

Application
Recommended Usage

Immunohistochemistry

5-15 µg/mL
Sample: Immersion fixed paraffin-embedded sections of human breast cancer tissue and immersion fixed paraffin-embedded sections of human liver cancer tissue

Immunoprecipitation

25 µg/mL
Sample: Conditioned cell culture medium spiked with Recombinant Human Serpin E1/PAI‑1 (Catalog # 1786-PI), see our available Western blot detection antibodies

Simple Western

2.5 µg/mL
Sample: HUVEC human umbilical vein endothelial cells

Western Blot

1 µg/mL
Sample: HUVEC human umbilical vein endothelial cells

Neutralization

Measured by its ability to neutralize Recombinant Human Serpin E1 (1.35 µg/mL, Catalog # 1786-PI) inhibition of Recombinant Human u‑Plasminogen Activator (uPA)/Urokinase (0.1 µg/mL, Catalog # 1310-SE) cleavage of the fluorogenic peptide substrate Z-GGR-AMC (100 µM). The Neutralization Dose (ND50) is typically 10 µg/mL.

Reviewed Applications

Read 1 review rated 4 using AF1786 in the following applications:

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: Serpin E1/PAI-1

As a member of the Serpin superfamily of serine protease inhibitors, Serpin E1/PAI-1 is the principal inhibitor of urokinase-type plasminogen activator (uPA) and tissue-type PA (1, 2). As important regulators of extracellular matrix remodeling, uPA and PAI-1 play a major role in many processes such as angiogenesis, tumor invasion and obesity (2-4). For example, uPA and PAI-1 are the only tumor prognostic factors validated at the highest level of evidence with regard to their clinical utility in breast cancer (5). The human PAI-1 is initially synthesized as 402 amino acid precursor with a N-terminal signal peptide (6, 7). PAI-1 may exist in one of two possible conformations, designated as active or latent (8). The purified recombinant human (rh) PAI-1 is active against rhuPA. The heterogeneity at the N-terminus of the purified rhPAI-1 has been observed before for both the recombinant and native proteins (9).

References

  1. Silverman, G.A. et al. (2001) J. Biol. Chem. 276:33293.
  2. Stefansson, S. et al. (2003) Curr. Pharm. Des. 9:1545.
  3. Duffy, M.J. (2002) Clin. Chem. 48:1194.
  4. Juhan-Vague, I. et al. (2003) J. Thromb. Haemost. 1:1575.
  5. Harbeck, N. et al. (2002) Clin. Breast Cancer 3:196.
  6. Pannekoek, H. et al. (1986) EMBO J. 5:2539.
  7. Ginsburg, D. et al. (1986) J. Clin. Invest. 78:1673.
  8. Wang, Z. et al. (1996) Biochemistry 35:16443.
  9. Stromqvist, M. et al. (1994) Protein Expr. Purif. 5:309.  

Long Name

Plasminogen Activator Inhibitor

Alternate Names

Nexin, PAI-1, PAI1, PLANH1

Entrez Gene IDs

5054 (Human); 18787 (Mouse)

Gene Symbol

SERPINE1

UniProt

P05121

Additional Serpin E1/PAI-1 Products

Product Documents for Human Serpin E1/PAI-1 Antibody

Certificate of Analysis

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Note: Certificate of Analysis not available for kit components.

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Product Specific Notices for Human Serpin E1/PAI-1 Antibody

For research use only

Citations for Human Serpin E1/PAI-1 Antibody

Customer Reviews for Human Serpin E1/PAI-1 Antibody (1)

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  • Human Serpin E1/PAI-1 Antibody
    Name: Anonymous
    Application: ELISA
    Sample Tested: Serum
    Species: Human
    Verified Customer | Posted 11/07/2018
    Combined with MAB1786 this antibody was used to make an immunoassay that was able to detect 3pg/ml of PAI-1.
    Human Serpin E1/PAI-1 Antibody AF1786

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Protocols

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Associated Pathways

Blood Coagulation Signaling Pathways
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