ANTI-APAF-1 CLONE 2E12

General description

Apoptotic protease-activating factor 1 (UniProt: O14727; also known as Apaf-1) is encoded by the Apaf-1 gene (Gene ID: 317) in human. Apaf-1 contains a caspase recruitment domain (CARD), an ATPase domain (NB-ARC), few short helical domains and then several copies of the WD40 repeat domain. It is highly expressed in lung and spleen, weakly in brain and kidney and is not detectable in liver. Under basal conditions it is a monomeric protein, but oligomerizes under apoptotic conditions. The monomeric form is auto-inhibited in a closed conformation through a bound ADP at the nucleotide binding site. Under apoptosis signals ADP is exchanged for ATP and its binding to cytochrome c triggers a conformation change where WD repeat region swings out, which allows the NB-ARC domain to rotate and expose the contact areas for oligomerization. Oligomeric form is a heptameric ring that binds to cytochrome c and ATP and is known as the apoptosome. Oligomeric Apaf-1 mediates the cytochrome c-dependent autocatalytic activation of pro-caspase-9 to mature caspase-9, which then activates effector caspases 3, 6, and 7.

Specificity

Clone 2E12 detects Apaf-1 in Jurkat cells. It targets an epitope within first 97 amino acids from the N-terminal region.

Immunogen

Epitope: N-terminus

Recombinant human Apaf-1 (aa 1-464) containing the N-terminal CARD and CED-4 homologous domains.

Application

Research Category
Apoptosis & Cancer

This rat monoclonal Anti-Apaf-1 Antibody, clone 2E12, Cat. No. MAB3503-I, has been tested for use in Electron Microscopy, Immunocytochemistry, Immunoprecipitation, and Western Blotting for the detection Apaf-1.

Western Blotting Analysis: A representative lot detected Apaf-1 in Western Blotting applications (Hausmann, G., et. al. (2000). J Cell Biol. 149(3):623-34).

Western Blotting Analysis: A representative lot detected Apaf-1 in Western Blotting applications (Moriishi, K., et. al. (1999). Proc Natl Acad Sci USA. 96(17):9683-8).

Immunocytochemistry Analysis: A representative lot detected Apaf-1 in Immunocytochemistry applications (Hausmann, G., et. al. (2000). J Cell Biol. 149(3):623-34).

Immunoprecipitation Analysis: A representative lot detected Apaf-1 in Immunoprecipitation applications (Moriishi, K., et. al. (1999). Proc Natl Acad Sci USA. 96(17):9683-8).

Electron Microscopy Analysis: A representative lot detected Apaf-1 in Electron Microscopy applications (Hausmann, G., et. al. (2000). J Cell Biol. 149(3):623-34).

Quality

Evaluated by Western Blotting in Jurkat cell lysate.

Western Blotting Analysis: 1 µg/mL of this antibody detected Apaf-1 in 10 µg of Jurkat cell lysate.

Target description

~140 kDa observed; 141.84 kDa calculated. Uncharacterized bands may be observed in some lysate(s).

Physical form

Format: Purified

Protein G purified

Purified rat monoclonal antibody IgG2a in PBS with 0.02% sodium azide.

Storage and Stability

Stable for 1 year at 2-8°C from date of receipt.

Other Notes

Concentration: Please refer to lot specific datasheet.

Disclaimer

Unless otherwise stated in our catalog or other company documentation accompanying the product(s), our products are intended for research use only and are not to be used for any other purpose, which includes but is not limited to, unauthorized commercial uses, in vitro diagnostic uses, ex vivo or in vivo therapeutic uses or any type of consumption or application to humans or animals.