APE (Ref-1, Apurinic Apyrimidinic Endonuclease, APEX, APE-1, HAP1)

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.\n\nApplications:\nSuitable 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. \n\nRecommended Dilution: \nWestern 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.\nImmunohistochemistry: 1:100-1:200 on frozen and paraffin sections from human and rat tissues. \nImmunocytochemistry: 1:100-1:200 on cytospin preparations of cells fixed in paraformaldehyde.\nApoptosis (TUNEL) assay: perform antibody staining AFTER the \nTUNEL assay.\nOptimal dilutions to be determined by the researcher. \n\nStorage and Stability:\nMay be stored at 4 degrees C for short-term only. For long-term storage and to avoid repeated freezing and thawing, add sterile glycerol (40-50%), aliquot and store at -20 degrees C. Aliquots are stable for at least 12 months at -20 degrees C. 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.

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SPECIFICATIONS

Catalog Number

A2298-74K

Size

100ul

Applications

ICC, IHC, WB

Hosts

Rabbit

Reactivities

Hum, Mouse, Rat

Form

Supplied as a liquid in PBS. No preservative added.

P Type

Pab

Purity

Purified by immunoaffinity chromatography.

Isotype

IgG

References

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) \n2. Focal nuclear hepatocyte response to oxidative damage following low dose thioacetamide intoxication: G.A. Clawson, et al.; Carcinogenesis 18, 1663 (1997) \n3. 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) \n4. 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)

Additional Info

Recognizes human APE. Detects a band of ~37kD by Western Blot\nSpecies Crossreactivity: mouse and rat