Technical Data
A2298-74K
APE (Ref-1, Apurinic Apyrimidinic Endonuclease, APEX, APE-1, HAP1)
Description:
Mammalian apurinic/apyrimidinic (AP) endonuclease (APE1; APEX; HAP1) is a multifunctional, bipartite enzyme that belongs to the class II AP endonucleases and that plays an important role in numerous, cellular functions, like repairing abasic sites (loss of purine or pyrimidine base) in DNA, regulating the redox state of other proteins that play roles in oxidative signalling, transcription factor regulation (Fos, Jun, NF-.kappa.B, Myb, HIF-1 alpha, CREB, Pac), cell cycle control (p53) and apoptosis. APE1 initiates the DNA base excision repair (BER) by cleaving the DNA immediately adjacent to the 5' of an AP site to produce a hydroxyl group at the 3' terminus of an unmodified nucleotide upstream of the nick and a 5'-deoxyribose phosphate moiety downstream. The product of APE1 is further processed by DNA polymerase beta to release 5'-deoxyribose phosphate with its intrinsic lyase activity. The nicked DNA is then sealed by DNA ligase I or DNA ligaseIII/XRCC1 to complete this repair process. In addition to the AP endonuclease activity, APE1 also possesses a 3'-5' DNA exonuclease activity, a 3'-phosphodiesterase activity, a 3'-phosphatase activity, and a RNase H activity. Recently it has been shown that APE1 is capable of excision L-configuration deoxyribonucleoside analogs from the 3' termini of DNA. Human APE1 gene is located on chromosome 14q 11.2-12.

Applications:
Suitable for use in Immunocytochemistry, Western Blot and Immunohistochemistry. Also suitable for use in Apoptosis assay (following TUNEL assay using Boehriner-Mannheim TUNEL assay kit). Other applications not tested.

Recommended Dilution:
Western Blot: 1:300-1:1000 on HL60 and HeLa cells using ECL. Antibody may be used 1-4X in the binding solution depending on the cells/cell lines.
Immunohistochemistry: 1:100-1:200 on frozen and paraffin sections from human and rat tissues.
Immunocytochemistry: 1:100-1:200 on cytospin preparations of cells fixed in paraformaldehyde.
Apoptosis (TUNEL) assay: perform antibody staining AFTER the
TUNEL assay.
Optimal dilutions to be determined by the researcher.

Storage and Stability:
May be stored at 4C for short-term only. For long-term storage and to avoid repeated freezing and thawing, add sterile glycerol (40-50%), aliquot and store at -20C. Aliquots are stable for at least 12 months at -20C. For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap. Further dilutions can be made in assay buffer.
TypeIsotypeCloneGrade
PabIgGAffinity Purified
SizeStorageShippingSourceHost
100ul4C (-20C Glycerol)Blue IceHumanRabbit
Concentration:
~0.1mg/ml
Immunogen:
Recombinant human APE
Purity:
Purified by immunoaffinity chromatography.
Form
Supplied as a liquid in PBS. No preservative added.
Specificity:
Recognizes human APE. Detects a band of ~37kD by Western Blot Species Crossreactivity: mouse and rat
Intended for research use only. Not for use in human, therapeutic, or diagnostic applications.
1. The apurinic/apyrimidinic endonuclease (APE/ref-1) DNA repair enzyme is elevated in premalignant and malignant cervical cancer: Y. Xu, et al.; Anticancer Res. 17, 3713 (1997)
2. Focal nuclear hepatocyte response to oxidative damage following low dose thioacetamide intoxication: G.A. Clawson, et al.; Carcinogenesis 18, 1663 (1997)
3. Down-regulation of apurinic/apyrimidinic endonuclease expression is associated with the induction of apoptosis in differentiating myeloid leukemia cells: K.A. Robertson, et al.; Cell Growth Differentiation 8, 443 (1997)
4. Differential cellular and subcellular expression of the human multifunctional apurinic/apyrimidinic endonuclease (APE/ref-1) DNA repair enzyme: J.R. Duguid, et al.; Cancer Res. 55, 6097 (1995)