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

The mammalian apurinic/apyrimidinic endonuclease (APE/ref-1) is responsible for the repair of AP sites in DNA. In addition, this enzyme has been shown to function as a redox factor facilitating the DNA binding capability of FOS, JUN, NfkB, HIF-1a as well as other transcription factors. Recently, APE has also been shown to control p53 activity through redox alteration. APE is also linked to apoptosis, associated with thioredoxin, and altered levels of APE have been found in some cancers (3). APE appears to form a unique link between the DNA base excision pathway, oxidative signaling, transcription regulation, cancer and cell-cycle control (4).\n\nApplications:\nSuitable for use in Western Blot, Immunohistochemistry and Apoptosis. Other applications not tested.\n\nRecommended Dilution:\nWestern Blot: 1:500 (ECL). Detects a single band at 37kD. Diluted antibody can be reused 1-3 times in Western Blot analysis.\nImmunohistochemistry (Frozen, fixed paraffin, cytospin preps): 1:50.\nApoptosis: Can be used following the apoptosis (TUNEL) procedure using the Boehringer-Mannheim TUNEL assay kit (2). Antibody staining should be performed AFTER the TUNEL assay.\nOptimal dilutions to be determined by researcher.\n\nPositive Control: HeLa whole cell extract.\n\nCellular Localization: \nIn Immunohistochemistry, punctate nuclear staining is seen in most cell types. In hepatocytes, large neurons and granule cells, cytoplasmic staining is observed.\n\nStorage and Stability:\nMay be stored at 4 degrees C for short-term only. For long-term storage, 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

A2298-74A

Size

100ul

Applications

IHC, WB

Hosts

Rabbit

Reactivities

Hum

Form

Supplied as a liquid in PBS, pH 7.2, 0.05% sodium azide, before the addition of glycerol to 40%.

P Type

Pab

Purity

Purified by immunoaffinity chromatography.

Isotype

IgG

References

1. Duguid, J. R., Eble, J. N., Wilson, T. M., and Kelley, M. R. Differential cellular and subcellular expression of the human multifunctional apurinic/apyrimidinic endonuclease (APE/ref-1) DNA repair enzyme, Cancer Res. 55: 6097-102, 1995. 2. Robertson, K. A., et al, Downregulation of AP endonuclease expression is associated with the induction of apoptosis in differentiating myeloid leukemia cells., Cell Growth Different. 8: 443-449, 1997. 3. Xu, Y., et al, The apurinic/apyrimidinic (Ape/ref-1) DNA repair enzyme is elevated in premalignant and malignant cervical cancer, Anticancer Res.,7(5):3713-3720,1997. 4. Xanthoudakis, S. and Curran, T. Redox Regulation of AP-1: A Link between Transcription Factor Signaling and DNA Repair. Adv. Expt. Med. Biol.,387: 69-75, 1996.\n5. Kelley, M.R., et al. The multifunctional DNA base excision repair and redox protein, AP endonuclease (APE/ref-1), and its role in germ cell tumors. Germ Cell Tumors IV. J. Libbey & Co., London,UK, 81-86, 1998.

Additional Info

Specific to the human APE/ref-1protein. Competitively inhibited from recognizing the APE/ref-1 antigen in tussues using APE/ref-1 prptein. Inhibits repair activity of ape protein.