APAF1 anticorps (N-Term)

N° du produit ABIN5540752

Cet anticorps anti-APAF1 est un anticorps Souris Monoclonal détectant APAF1 dans WB. Adapté pour Humain, Souris et Rat.

Aperçu rapide pour APAF1 anticorps (N-Term) (ABIN5540752)

Antigène: APAF1 (Apoptotic Peptidase Activating Factor 1 (APAF1))

Reactivité: Humain, Souris, Rat

Hôte: Souris

Clonalité: Monoclonal

Conjugué: Cet anticorp APAF1 est non-conjugé

Application: Western Blotting (WB)

Clone: 5C1

Détail du produit anti-APAF1 anticorps

Épitope: AA 1-421, N-Term

Specificité: This antibody detects ~130 kDa of human Apaf-1 protein from whole cell lysate of human cell lines.

Purification: Protein-A Sepharose

Immunogène: His-tagged human Apaf-1 fusion protein corresponding to N-terminal amino acids (1-421 aa)

Isotype: IgG1

Information d'application

Indications d'application: Western blot: 5 μg/mL. For details see protocol below.

Protocole: SDS PAGE & Western Blotting 1) Rinse 1 x 10e7 of cells 3 times with PBS and suspend it with 1 mL of 1 x Laemmli SDS-PAGE sample buffer, then lyse the cells by brief sonication (up to 20 seconds). 2) Boil the sample for 3-5 minutes and centrifuge at 12,000 x g for a minute. Other methods may be suitable. 3) Resolve 20 μ l of the sample per lane by 7.5 % SDS- polyacrylamide gel electrophoresis (See the manufacture's manual for electrophoresis conditions). 4) Blot the protein to a polyvinylidene difluoride (PVDF) membrane at 1 mA/cm 2 for 1 hour in a semi-dry transfer system. (Transfer Buffer: 25 mM Tris, 190 mM glycine, 20 % Methanol). See the manufacture's manual for precise transfer procedure. 5) To reduce nonspecific binding, soak the membrane in Block Ace TM (Snow bland) for 1 hour at 37 o C, or overnight at 4 o C. 6) Dilute the primary antibody to 5 μ g/mL with Dilution Buffer (10 % Block Ace TM /PBS). Incubate the membrane with primary antibody dilution for 1 hour at room temperature (20-25 o C). (The optimal concentration of antibody will depend on the experimental conditions.) 7) Wash the membrane with Wash Buffer (0.2 % Tween20 in PBS) for 5-10 minutes x 3 times. 8) Incubate the membrane with secondary antibody (1:10,000 diluted horseradish peroxidase conjugated anti Mouse IgG goat antibody in Dilution Buffer) for 1 hour at room temperature. 9) Wash the membrane with Wash buffer (10 minutes x 3 times) 10) Wipe excess buffer from the membrane, then incubate it with appropriate chemiluminescence reagents for 1 minute. Remove excess reagent from the membrane by dabbing with paper towel, and seal it in plastic wrap. 11) Expose to X-ray film in a dark room for 30 seconds. Develop the film as usual. The conditions for exposure and development may vary.

Restrictions: For Research Use only

Stockage

Format: Liquid

Buffer: PBS containing 50 % glycerol. Contains no preservatives.

Agent conservateur: Without preservative

Stock: -20 °C

Stockage commentaire: Upon receipt, store undiluted (in aliquots) at -20°C. Avoid repeated freezing and thawing. Shelf life: One year from despatch.

Détails sur APAF1

Antigène: APAF1 (Apoptotic Peptidase Activating Factor 1 (APAF1))

Autre désignation: apaf1

Sujet: Apoptosis occurs during normal cellular development and involves dramatic changes in cellular structure. Disruption of apoptosis may contribute to cancer as well as other autoimmune diseases. Recently, the mammalian homologue of the key cell death gene CED-4 in C. elegans has been identified from human and mouse, and designated Apaf-1 (apoptosis protease activating factor 1) (Ref. 1,2). Apaf-1 binds to cytochrome c (Apaf-2), a reaction that requires dATP, and this leads to the formation of a large multimeric complex. The complex recruits pro-caspase-9, and activates caspase-9 (Apaf-3). Activated caspase-9 in turn cleaves and activates caspase-3, one of the proteases responsible for the proteolytic cleavage of many key proteins in apoptosis. Apaf-1 is ubiquitously expressed in human tissues (1).

UniProt: O14727

Pathways: Signalisation p53, Apoptose, Caspase Cascade in Apoptosis, Tube Formation, Positive Regulation of Endopeptidase Activity