APE Antibody (12H1)

Western Blot: APE Antibody (12H1) [NBP2-76919] - Western blot analysis of APE1 on HL-60 lysates. Proteins were transferred to a PVDF membrane and blocked with 5% BSA in PBS for 1 hour at room temperature. The primary antibody was used at a 1:1,000 dilution in 5% BSA at room temperature for 2 hours. Goat Anti-Mouse IgG - HRP Secondary Antibody (HA1006) at 1:5,000 dilution was used for 1 hour at room temperature.

Immunocytochemistry/Immunofluorescence: APE Antibody (12H1) [NBP2-76919] - ICC staining APE1 in SiHa cells (green). Formalin fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 10 minutes at room temperature and blocked with 1% Blocker BSA for 15 minutes at room temperature. Cells were probed with APE1 monoclonal antibody at a dilution of 1:50 for 1 hour at room temperature, washed with PBS. Alexa Fluorc 488 Goat anti-Mouse IgG was used as the secondary antibody at 1/100 dilution. The nuclear counter stain is DAPI (blue).

Immunohistochemistry: APE Antibody (12H1) [NBP2-76919] - Immunohistochemical analysis of paraffin-embedded mouse colon tissue using anti-APE1 antibody. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) for 20 minutes. The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the antibody at 1/200 dilution, for 30 minutes at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chrogen. Counter stained with hematoxylin and mounted with DPX.

Flow Cytometry: APE Antibody (12H1) [NBP2-76919] - Flow cytometric analysis of APE1 was done on SiHa cells. The cells were fixed, permeabilized and stained with APE1 antibody at 1/100 dilution (red) compared with an unlabelled control (cells without incubation with primary antibody; black). After incubation of the primary antibody on room temperature for an hour, the cells was stained with a Alexa Fluor 488-conjugated goat anti-mouse IgG Secondary antibody at 1/500 dilution for 30 minutes.

Immunocytochemistry/Immunofluorescence: APE Antibody (12H1) [NBP2-76919] - ICC staining APE1 in A549 cells (green). Formalin fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 10 minutes at room temperature and blocked with 1% Blocker BSA for 15 minutes at room temperature. Cells were probed with APE1 monoclonal antibody at a dilution of 1:50 for 1 hour at room temperature, washed with PBS. Alexa Fluorc 488 Goat anti-Mouse IgG was used as the secondary antibody at 1/100 dilution. The nuclear counter stain is DAPI (blue).

Immunohistochemistry: APE Antibody (12H1) [NBP2-76919] - Immunohistochemical analysis of paraffin-embedded human liver tissue using anti-APE1 antibody. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) for 20 minutes. The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the antibody at 1/200 dilution, for 30 minutes at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chrogen. Counter stained with hematoxylin and mounted with DPX.

Immunohistochemistry: APE Antibody (12H1) [NBP2-76919] - Immunohistochemical analysis of paraffin-embedded human prostate cancer tissue using anti-APE1 antibody. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) for 20 minutes. The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the antibody at 1/200 dilution, for 30 minutes at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chrogen. Counter stained with hematoxylin and mounted with DPX.

Immunohistochemistry: APE Antibody (12H1) [NBP2-76919] - Immunohistochemical analysis of paraffin-embedded human kidney tissue using anti-APE1 antibody. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) for 20 minutes. The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the antibody at 1/200 dilution, for 30 minutes at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chrogen. Counter stained with hematoxylin and mounted with DPX.

Product Discontinued

APE Antibody (12H1) Summary

• Immunogen: Recombinant protein within Human APE aa 20-350. (SwissProt: P27695 Human; SwissProt: P28352 Mouse)
• Localization: Nucleus. Endoplasmic reticulum.
• Isotype: IgG2a
• Clonality: Monoclonal
• Host: Mouse
• Gene: APEX1
• Purity: Immunogen affinity purified

Applications/Dilutions

• Dilutions:
  • Flow Cytometry 1:50-1:100
  • Immunocytochemistry/ Immunofluorescence 1:50
  • Immunohistochemistry
  • Immunohistochemistry-Paraffin 1:50-1:200
  • Western Blot 1:1000-1:5000
• Theoretical MW: 36 kDa.
  Disclaimer note: The observed molecular weight of the protein may vary from the listed predicted molecular weight due to post translational modifications, post translation cleavages, relative charges, and other experimental factors.

Reactivity Notes

Mouse blocking reagent may be needed for IHC and ICC experiments to reduce high background signal. You can find these reagents under catalog numbers PK-2200-NB and MP-2400-NB. Please contact Technical Support if you have any. Please note that this antibody is reactive to Mouse and derived from the same host, Mouse. Mouse-On-questions.

Packaging, Storage & Formulations

• Storage: Store at 4C short term. Aliquot and store at -20C long term. Avoid freeze-thaw cycles.
• Buffer: PBS (pH7.4), 0.2% BSA, 50% Glycerol
• Preservative: 0.05% Sodium Azide
• Concentration: 2 mg/ml
• Purity: Immunogen affinity purified

Alternate Names for APE Antibody (12H1)

• AP endonuclease 1
• APE
• APE1
• APE-1
• APEAPEX nuclease (multifunctional DNA repair enzyme)
• APENAP endonuclease class I
• APEX deoxyribonuclease (apurinic or apyrimidinic)
• APEX nuclease (multifunctional DNA repair enzyme) 1
• APEX nuclease
• APEX1
• Apurinic-apyrimidinic endonuclease 1
• APXAP lyase
• DNA-(apurinic or apyrimidinic site) lyase
• EC 3.1
• EC 4.2.99.18
• HAP1apurinic/apyrimidinic (abasic) endonuclease
• multifunctional DNA repair enzyme
• protein REF-1
• redox factor 1
• Redox factor-1
• REF1 apurinic/apyrimidinic exonuclease
• REF-1

Background

DNA-(apurinic or apyrimidinic site) lyase, also known as APE1 or APEX1, is a multifunctional mammalian proteins responsible for the cellular response to oxidative stress. Specifically, APE-1 both functions in the DNA base excision repair pathway and regulates the redox of transcriptional factors.

For DNA repair, APE1 functions as a mammalian apurinic/apyrimidinic (AP) endodeoxyribonuclease responsible for the repair of AP sites in DNA lesions induced by oxidative and alkylating agents.

APEX1 also exerts a reversible nuclear redox activity to regulate the DNA binding affinity and the activity of transcriptional factors by regulating the redox status of their DNA-binding domains.

APE1 antibodies are useful for studies on both DNA repair and transcriptional regulation.

Limitations

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.

Bioinformatics

• Gene Symbol: APEX1