Anti-APE/Ref-1 Antibody, clone 13B8E5C2

DNA-(apurinic or apyrimidinic site) lyase (EC 4.2.99.18; UniProt P27695; also known as AP endonuclease 1, APE-1, APEN, APEX nuclease, Apurinic-apyrimidinic endonuclease 1, Redox factor-1, REF-1) is encoded by the APEX1 (also known as APE, APE1, APEX, APX, HAP1, REF1) gene (Gene ID 328) in human. APE/Ref-1 mediates the cleavage of an apurinic/apyrimidinic (AP) site in DNA generated by DNA glycosylases during base excision repair (BER) in both nucleus and mitochondria. APE/Ref-1 also functions as a redox factor (ref) responsible for maintaining several transcription factors (TFs) in an active reduced state for DNA binding, including AP-1, NF-κB, HIF-1α, CREB, TP53. Elevated levels of APE1 are associated with resistance to chemotherapyand poor prognosis in various cancers. APE/Ref-1 is synthesized as a 318-a.a. protein whose initiation methionine (Met1) is removed posttranslationally to yield the 317-a.a. form with a nuclear localization sequence (NLS; a.a. 8-13) and a nuclear export signal (NES; a.a. 64-80) sequence to allow its shuttle between the cytoplasm and nucleus. The 317-a.a. form can be further processed by a mitochondria-associated N-terminal peptidase into a mitochrondrial form (mtAPE; a.a. 32-318) that lacks NLS and contains only the NES and the mitochondrial targeting sequence (MTS; a.a. 289-318).

Clone 13B8E5C2 detected both cytosolic and nuclear APE/Ref-1 by Immunocytochemistry, Immunohistochemistry, and Western blotting (Moore, D.H., et al. (2000). Clin. Cancer Res. 6(2):602-609).

Recombinant full-length human APE/Ref-1.

Anti-APE/Ref-1, clone 13B8E5C2, Cat. No. MABE1068, is a highly specific mouse monoclonal antibody, that targets APE1 and has been tested in Immunocytochemistry, Immunohistochemistry (Paraffin), and Western Blotting.

Immunocytochemistry Analysis: A 1:200 dilution from a representative lot detected both cytosolic and nuclear APE/Ref-1 immunoreactivity in 4% paraformaldehyde-fixed, 0.3% Triton X-100-permeabilized HeLa cells.

Immunohistochemistry Analysis: A 1:1,000 dilution from a representative lot detected nuclear APE/Ref-1 immunoreactivity in rat kidney, and human placenta & small intestine tissue sections.

Western Blotting Analysis: 0.5 µg/mL from a representative lot detected APE/Ref-1 in 10 µg of Raji cell lysate.

Immunocytochemistry Analysis: A representative lot detected both cytosolic and nuclear APE/Ref-1 immunoreactivity in HeyC2 and HEY human ovarian cancer cell lines (Moore, D.H., et al. (2000). Clin. Cancer Res. 6(2):602-609).

Immunohistochemistry Analysis: A representative lot detected varying tissue distribution and cellular localization of APE/Ref-1 immunoreactivity among various 4% formaldehyde-fixed, paraffin-embedded human ovarian tissue samples, including ovarian endometriosis, serous cystadenoma , serous carcinoma, ands well as normal ovarian tissues (Moore, D.H., et al. (2000). Clin. Cancer Res. 6(2):602-609).

Western Blotting Analysis: A representative lot detected an upregulated cytosolic APE/Ref-1 level in HeyC2 than HEY cells, while similar nuclear APE/Ref-1 level was detected in these two human ovarian cancer cell lines (Moore, D.H., et al. (2000). Clin. Cancer Res. 6(2):602-609).

Evaluated by Western Blotting in HepG2 cell lysate.

Western Blotting Analysis: 0.5 µg/mL of this antibody detected APE/Ref-1 in 10 µg of HepG2 cell lysate.

~35-39 kDa observed. 35.42 kDa (Met1 removed; a.a. 2-318) and 32.24 kDa (Mitochondrial form; a.a. 32-318) calculated. The broad banding pattern is consistent with the detection of both mitochondrial and non-mitochondrial forms with varying degrees of posttranslational modifications (lysine acetylation, threonine phosphorylation, cysteine nitrosylation). Uncharacterized bands may be observed in some lysate(s).

Format: Purified

Concentration: Please refer to lot specific datasheet.