APE Antibody

Novus Biologicals | Catalog # NB100-897

Novus Biologicals
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Key Product Details

Species Reactivity

Validated:

Human

Cited:

Human

Predicted:

Bovine (100%), Canine (100%), Porcine (100%). Backed by our 100% Guarantee.

Applications

Validated:

Immunohistochemistry, Immunohistochemistry-Paraffin, Western Blot, Peptide ELISA, Immunocytochemistry/ Immunofluorescence

Cited:

Western Blot, Immunocytochemistry/ Immunofluorescence

Label

Unconjugated

Antibody Source

Polyclonal Goat IgG
View all APE Primary Antibodies »

Product Specifications

Immunogen

Peptide with sequence PKRGKKGAVAEDGD-C corresponding to N-Terminus according to NP_001632.2.

Reactivity Notes

Use in Human reported in scientific literature (PMID:33753167).

Specificity

Reported variants represent identical protein (NP_001632.2; NP_542379.1; NP_542380.1).

Clonality

Polyclonal

Host

Goat

Isotype

IgG

Description

Novus Biologicals Goat APE Antibody (NB100-897) is a polyclonal antibody validated for use in IHC, WB, ELISA and ICC/IF. Anti-APE Antibody: Cited in 6 publications. All Novus Biologicals antibodies are covered by our 100% guarantee.

Scientific Data Images for APE Antibody

Western Blot: APE Antibody [NB100-897]

Western Blot: APE Antibody [NB100-897]

Western Blot: APE Antibody [NB100-897] - Staining of A431 (A), HeLa (B) and MCF7 (C) nuclear lysates (35ug protein in RIPA buffer). Primary incubation was 1 hour. Detected by chemiluminescence.
Immunohistochemistry-Paraffin: APE Antibody [NB100-897]

Immunohistochemistry-Paraffin: APE Antibody [NB100-897]

Immunohistochemistry-Paraffin: APE Antibody [NB100-897] - Staining of paraffin embedded Human Breast. Steamed antigen retrieval with Tris/EDTA buffer pH 9, HRP-staining.
APE Antibody

Western Blot: APE Antibody [NB100-897] -

APE1 EVs secretion in a panel of mammalian cell lines:A, western blotting analyses for APE1 detection carried out in EXE of JHH-6, A549, HCT116P53+/+, HCT116p53-/-, CH13F3 delta /+/+, and CH12F3 delta / delta / delta cells. Both APE1 p37 and p33 forms have been detected. Alix was used as a loading control. WCE were analyzed as a positive control of APE1 expression. The molecular mass (MW) expressed in kDa is shown on the left side of the image. B, western blotting analyses for APE1 detection carried out in EXE of MEFNPM1+/+ and MEFNPM1-/- cells. Both APE1 p37 and p33 forms have been detected. Alix was used as a loading control. WCE were analyzed as a positive control of APE1 expression and beta -Tubulin (apparent molecular weight 52 kDa) was detected as loading control. Western blotting analysis for NPM1 detection was carried out as a control for cells genotype. The molecular mass (MW) expressed in kDa is shown on the left side of the image. C, western blotting analyses for APE1 detection in WCE and EXE derived from OCI-AML2 and OCI-AML3 cell lines. beta -Tubulin (apparent molecular weight 52 kDa) and Alix (apparent molecular weight 95 kDa) detection was carried out as loading controls. APE1 p37 and p33 are indicated by arrows. The molecular mass (MW) expressed in kDa is shown on the left side of the image. The percentages of exosomal APE1 p37 and p33 forms in OCI-AML2 and OCI-AML3 cell lines are indicated in the scatter plot on the right panel and in Fig. S5. Data are expressed as mean +/- SD of three independent replicas. Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/33753167), licensed under a CC-BY license. Not internally tested by Novus Biologicals.
APE Antibody

Western Blot: APE Antibody [NB100-897] -

APE1 EVs secretion in a panel of mammalian cell lines:A, western blotting analyses for APE1 detection carried out in EXE of JHH-6, A549, HCT116P53+/+, HCT116p53-/-, CH13F3 delta /+/+, and CH12F3 delta / delta / delta cells. Both APE1 p37 and p33 forms have been detected. Alix was used as a loading control. WCE were analyzed as a positive control of APE1 expression. The molecular mass (MW) expressed in kDa is shown on the left side of the image. B, western blotting analyses for APE1 detection carried out in EXE of MEFNPM1+/+ and MEFNPM1-/- cells. Both APE1 p37 and p33 forms have been detected. Alix was used as a loading control. WCE were analyzed as a positive control of APE1 expression and beta -Tubulin (apparent molecular weight 52 kDa) was detected as loading control. Western blotting analysis for NPM1 detection was carried out as a control for cells genotype. The molecular mass (MW) expressed in kDa is shown on the left side of the image. C, western blotting analyses for APE1 detection in WCE and EXE derived from OCI-AML2 and OCI-AML3 cell lines. beta -Tubulin (apparent molecular weight 52 kDa) and Alix (apparent molecular weight 95 kDa) detection was carried out as loading controls. APE1 p37 and p33 are indicated by arrows. The molecular mass (MW) expressed in kDa is shown on the left side of the image. The percentages of exosomal APE1 p37 and p33 forms in OCI-AML2 and OCI-AML3 cell lines are indicated in the scatter plot on the right panel and in Fig. S5. Data are expressed as mean +/- SD of three independent replicas. Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/33753167), licensed under a CC-BY license. Not internally tested by Novus Biologicals.
APE Antibody

Western Blot: APE Antibody [NB100-897] -

APE1 proteolytic activity characterization in exosomes:A, western blotting analyses for APE1 p37 and p33 detection in EVs derived from JHH-6 MG-132 treated cells and the respective untreated CTR cells (left panel). Alix (apparent molecular weight 95 kDa) was used as loading control. The percentage of exosomal APE1 p37 and p33 forms in both CTR and MG-132 JHH-6 cells has been obtained by densitometry analysis, and it is indicated in the scatter plot (middle panel) and in Fig. S6. Western blotting analysis for I kappa B-alpha (apparent molecular weight 38 kDa) detection was performed in CTR and MG-132 treated JHH-6 WCE as a confirmation of proteases inhibition. The molecular mass (MW), expressed in kDa, is shown on the left side of the image. Data are expressed as mean +/- SD of three independent replicas. B, EVs possess proteases able to cleave rGST-APE1 protein. In vitro proteolytic activity exerted by EXE (0.1 μg) upon rGST-APE1WT (0.2 μg) (apparent molecular weight 65 kDa). The reactions were performed at 37 C for the indicated times and the substrates (♦) and the products (♦♦) of reactions have been detected by Western blotting using APE1 antibody (left panel). rGST (0.2 μg) (apparent molecular weight 29 kDa) indicated as (•) has been detected by western blotting using GST antibody. C, graph illustrating the comparison of EXE cleavage activity upon rGST-APE1WT and rGST-APE1K4pleA. Data are expressed as mean +/- SD of three independent technical replicas. ∗p < 0.05, ∗∗p < 0.01, ∗∗∗p < 0.001, ∗∗∗∗p < 0.0001. Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/33753167), licensed under a CC-BY license. Not internally tested by Novus Biologicals.
APE Antibody

Western Blot: APE Antibody [NB100-897] -

EVs characterization and APE1 protein content analysis in JHH-6 cells:A, average size distribution curve determined by NTA of EVs derived from JHH-6 conditioned medium. The average concentration of vesicles was plotted against their size. The mean mode sizes and standard deviations are reported nearby the curve. B, western blotting analysis performed in JHH-6 WCE and EXE for APE1 detection (predicted molecular weight 37 kDa). APE1 p37 and p33 forms were observed in EXE. WCE were used as a positive APE1 control (lower panel). Golgi apparatus protein GM130 (apparent molecular weight 130 kDa) was assayed as exosome negative marker, Alix (apparent molecular weight 95 kDa) and Syntenin (apparent molecular weight 32 kDa) were assayed as exosome positive markers. C, AlphaScreen assay titrating the recombinant rGST-APE1 protein using Glutathione Donor beads and Protein A/G Acceptor beads in the presence of anti-APE1 antibody. The signal of the recombinant protein was detected in NBI-isolated vesicles upon a 95 C heating step. The specific fluorescence counts, indicating a high signal to background ratio, were normalized to background after subtraction of the aspecific binding when considered at two EV concentrations in the scatter plot. Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/33753167), licensed under a CC-BY license. Not internally tested by Novus Biologicals.
APE Antibody

Western Blot: APE Antibody [NB100-897] -

APE1 EVs secretion in a panel of mammalian cell lines:A, western blotting analyses for APE1 detection carried out in EXE of JHH-6, A549, HCT116P53+/+, HCT116p53-/-, CH13F3 delta /+/+, and CH12F3 delta / delta / delta cells. Both APE1 p37 and p33 forms have been detected. Alix was used as a loading control. WCE were analyzed as a positive control of APE1 expression. The molecular mass (MW) expressed in kDa is shown on the left side of the image. B, western blotting analyses for APE1 detection carried out in EXE of MEFNPM1+/+ and MEFNPM1-/- cells. Both APE1 p37 and p33 forms have been detected. Alix was used as a loading control. WCE were analyzed as a positive control of APE1 expression and beta -Tubulin (apparent molecular weight 52 kDa) was detected as loading control. Western blotting analysis for NPM1 detection was carried out as a control for cells genotype. The molecular mass (MW) expressed in kDa is shown on the left side of the image. C, western blotting analyses for APE1 detection in WCE and EXE derived from OCI-AML2 and OCI-AML3 cell lines. beta -Tubulin (apparent molecular weight 52 kDa) and Alix (apparent molecular weight 95 kDa) detection was carried out as loading controls. APE1 p37 and p33 are indicated by arrows. The molecular mass (MW) expressed in kDa is shown on the left side of the image. The percentages of exosomal APE1 p37 and p33 forms in OCI-AML2 and OCI-AML3 cell lines are indicated in the scatter plot on the right panel and in Fig. S5. Data are expressed as mean +/- SD of three independent replicas. Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/33753167), licensed under a CC-BY license. Not internally tested by Novus Biologicals.
APE Antibody

Western Blot: APE Antibody [NB100-897] -

APE1 proteolytic activity characterization in exosomes:A, western blotting analyses for APE1 p37 and p33 detection in EVs derived from JHH-6 MG-132 treated cells and the respective untreated CTR cells (left panel). Alix (apparent molecular weight 95 kDa) was used as loading control. The percentage of exosomal APE1 p37 and p33 forms in both CTR and MG-132 JHH-6 cells has been obtained by densitometry analysis, and it is indicated in the scatter plot (middle panel) and in Fig. S6. Western blotting analysis for I kappa B-alpha (apparent molecular weight 38 kDa) detection was performed in CTR and MG-132 treated JHH-6 WCE as a confirmation of proteases inhibition. The molecular mass (MW), expressed in kDa, is shown on the left side of the image. Data are expressed as mean +/- SD of three independent replicas. B, EVs possess proteases able to cleave rGST-APE1 protein. In vitro proteolytic activity exerted by EXE (0.1 μg) upon rGST-APE1WT (0.2 μg) (apparent molecular weight 65 kDa). The reactions were performed at 37 C for the indicated times and the substrates (♦) and the products (♦♦) of reactions have been detected by Western blotting using APE1 antibody (left panel). rGST (0.2 μg) (apparent molecular weight 29 kDa) indicated as (•) has been detected by western blotting using GST antibody. C, graph illustrating the comparison of EXE cleavage activity upon rGST-APE1WT and rGST-APE1K4pleA. Data are expressed as mean +/- SD of three independent technical replicas. ∗p < 0.05, ∗∗p < 0.01, ∗∗∗p < 0.001, ∗∗∗∗p < 0.0001. Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/33753167), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Applications for APE Antibody

Application
Recommended Usage

Immunohistochemistry

4 - 6 ug/ml

Immunohistochemistry-Paraffin

4 - 6 ug/ml

Peptide ELISA

Detection limit 1:16000

Western Blot

0.1 - 0.3 ug/ml
Application Notes
WB: Approx. 37 kDa band observed in nuclear lysates of A431, HeLa and MCF7 (calculated MW of 35.6 kDa band according to NP_001632.2, NP_542379.1 and NP_542380.1).Use in ICC/IF reported in scientific literature (PMID: 27560983).

Formulation, Preparation, and Storage

Purification

Immunogen affinity purified

Formulation

Tris saline (20 mM Tris pH 7.3, 150 mM NaCl), 0.5% BSA

Preservative

0.02% Sodium Azide

Concentration

0.5 mg/ml

Shipping

The product is shipped with polar packs. Upon receipt, store it immediately at the temperature recommended below.

Stability & Storage

Store at -20C. Avoid freeze-thaw cycles.

Background: APE

Apurinic / apyrimidinic endonuclease (APE1 or REF1) is responsible for the incision of DNA basic sites during base excision repair. It has also been shown to stimulate the DNA binding activity of numerous transcription factors that are involved in cancer promotion and progression. Alterations in APE1 levels have been described in some cancers. APE1 is part of the cellular response to oxidative stress and protects cells from the genotoxic and cytotoxic effect of oxidizing agents.

Long Name

Apurinic/Apyrimidinic Endonuclease

Alternate Names

APEX1

Entrez Gene IDs

328 (Human)

Gene Symbol

APEX1

UniProt

NP_001632 (Human)

Additional APE Products

Product Documents for APE Antibody

Certificate of Analysis

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Product Specific Notices for APE Antibody

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Primary Antibodies are guaranteed for 1 year from date of receipt.

Related Research Areas

Base Excision Repair

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