PRODUCT CODE: EM1801-13

APE1 Monoclonal Antibody (EM1801-13)

Applications

  • WB

  • IHC-P

  • FC

REACTIVITY

  • Human

  • Mouse

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.
  • 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.
  • 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 (EM1801-13) 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.
  • 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 (EM1801-13) 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.
  • 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 (EM1801-13) 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.
  • 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 (EM1801-13) 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 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.
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.

Applications

  • WB

  • IHC-P

  • FC

REACTIVITY

  • Human

  • Mouse

SPECIFICATIONS

Product Type

Mouse monoclonal primary

Product Name

APE1 Monoclonal Antibody (EM1801-13)

Immunogen

Recombinant protein within human ape1 aa 20-350.

Host

Mouse

Positive Control

HL-60, human liver tissue, human prostate cancer tissue, human kidney tissue, mouse colon tissue, SiHa.

Conjugation

Unconjugated

Clonality

Monoclonal

Clone Number

12H4

PROPERTIES

Form

Liquid

Storage Condition

Store at +4C after thawing. Aliquot store at -20C. Avoid repeated freeze / thaw cycles.

Storage Buffer

1*PBS (pH7.4), 0.2% BSA, 50% Glycerol. Preservative: 0.05% Sodium Azide.

Concentration

2 ug/ul

PURIFICATION

Protein affinity purified.

MOLECULAR WEIGHT

36 kDa

Isotype

IgG1

APPLICATION DILUTION

  • WB:1:1,000-1:5,000

  • IHC-P:1:50-1:200

  • FC:1:50-1:100

TARGET

UNIPROT #

PROTEIN NAME

APE1

SYNONYMS

AP endonuclease 1 antibody; AP endonuclease class I antibody; AP lyase antibody; APE 1 antibody; APE antibody; APE-1 antibody; APEN antibody; APEX 1 antibody; APEX antibody; APEX nuclease (multifunctional DNA repair enzyme) 1 antibody; Apex nuclease 1 antibody; APEX nuclease antibody; APEX1 antibody; APEX1_HUMAN antibody; Apurinic endonuclease antibody; Apurinic-apyrimidinic endonuclease 1 antibody; Apurinic/apyrimidinic (abasic) endonuclease antibody; Apurinic/apyrimidinic endonuclease 1 antibody; Apurinic/apyrimidinic exonuclease antibody; APX antibody; BAP1 antibody; Deoxyribonuclease (apurinic or apyrimidinic) antibody; DNA (apurinic or apyrimidinic site) lyase antibody; DNA-(apurinic or apyrimidinic site) lyase, mitochondrial antibody; EC 4.2.99.18 antibody; HAP 1 antibody; HAP1 antibody; Human Apurinic endonuclease 1 antibody; MGC139790 antibody; Multifunctional DNA repair enzyme antibody; Redox factor 1 antibody; Redox factor-1 antibody; REF 1 antibody; REF 1 protein antibody; REF-1 antibody; REF1 antibody; REF1 protein antibody

SEQUENCE SIMILARITIES

Belongs to the DNA repair enzymes AP/ExoA family.

POST-TRANSLATIONAL MODIFICATION

Phosphorylated. Phosphorylation by kinase PKC or casein kinase CK2 results in enhanced redox activity that stimulates binding of the FOS/JUN AP-1 complex to its cognate binding site. AP-endodeoxyribonuclease activity is not affected by CK2-mediated phosphorylation. Phosphorylation of Thr-233 by CDK5 reduces AP-endodeoxyribonuclease activity resulting in accumulation of DNA damage and contributing to neuronal death.; Acetylated on Lys-6 and Lys-7. Acetylation is increased by the transcriptional coactivator EP300 acetyltransferase, genotoxic agents like H(2)O(2) and methyl methanesulfonate (MMS). Acetylation increases its binding affinity to the negative calcium response element (nCaRE) DNA promoter. The acetylated form induces a stronger binding of YBX1 to the Y-box sequence in the MDR1 promoter than the unacetylated form. Deacetylated on lysines. Lys-6 and Lys-7 are deacetylated by SIRT1.; Cleaved at Lys-31 by granzyme A to create the mitochondrial form; leading in reduction of binding to DNA, AP endodeoxynuclease activity, redox activation of transcription factors and to enhanced cell death. Cleaved by granzyme K; leading to intracellular ROS accumulation and enhanced cell death after oxidative stress.; Cys-65 and Cys-93 are nitrosylated in response to nitric oxide (NO) and lead to the exposure of the nuclear export signal (NES).; Ubiquitinated by MDM2; leading to translocation to the cytoplasm and proteasomal degradation.

SUBCELLULAR LOCATION

Nucleus. Endoplasmic reticulum.

FUNCTION

The role of transcription factors in the regulation of gene expression is well established. Although the activity of these factors can be regulated by phosphorylation, evidence has indicated regulation of DNA binding mediated by changes in reduction-oxidation (redox) status. Mutational analysis has identified a single conserved cysteine residue mapping within the DNA binding domains of Fos and Jun. Chemical oxidation or modification of this cysteine residue inhibits the DNA binding activity of Fos and Jun. A similar mode of regulation has been recently proposed for other nuclear transcription factors. Oxidation is reversible by these compounds or by a cellular redox/DNA repair protein identified originally as Ref-1 (redox factor 1). Ref-1 is identical to a previously characterized DNA repair enzyme designated HAP1, APE or APEX.