Cytochrome C, Somatic (CYCS) Antikörper

Antigen: Cytochrome C, Somatic (CYCS)

Synonyme: cycs, CYCS, CYC, HCS, THC4, cyc, hcs, cyct, MGC75709, MGC89066, DKFZp468F1016, MGC128356, CYCSA, MGC93634, Cyt-c1, CytC-1, CytC1, Cytc, Cytc-d, DC3, bln, bs30f07.y1, cyt c-d, cyt-c-d, cyt.c, cytochrome c, ms(2)04445, pDMc02, DmelCG13263, CG13263, Cyt-c2, CytC-2, Cytc-p, DC4, DMc01, cyt c-p, cyt-c-p, l(2)SH1590, pDMc01, DmelCG17903, CG17903

Klonalität: Monoklonal (1E4)

Wirt: Huhn

Reaktivität: Human, Maus, Ratte (Rattus)

Applikation: Western Blot (WB)

Produktnummer: ABIN130465

Produktbeschreibung

Weitere Bezeichnung: Cytochrome C

Immunogen: This antibody was concentrated from hybridoma (clone 1E4) supernatant. This hybridoma was established by fusion of chicken B cell line MuH1 cell with chicken splenocyte immunized with the synthesized peptide,CKEERADLIAYLKKATNE which corresponds to the C-terminus of human cytochrome c.

Format: Supernatant

Isotyp: IgY  (Passende Sekundärantikörper)

Klon: 1E4

Beschreibung: Cytochrome c released from the mitochondria binds to Apaf-1 (Apoptotic protease activating factor-1). In the presence of dATP, the Apaf-1/cytochrome c complex undergoes a conformational change that exposes the caspase recruitment domain (CARD) of Apaf-1, allowing caspase-9 to bind. Activated caspase-9 in turn cleaves and activates caspase-3. Activated caspase-3 cleaves poly(adenosine diphosphate-ribose) polymerase (PARP), DNA-dependent protein kinase, nuclear lamins and several other cellular proteins, leading to apoptosis.

Spezifität: This antibody reacts with cytochrome c (15 kDa) on Western blots.

Synonyme: cycs, CYCS, CYC, HCS, THC4, cyc, hcs, cyct, MGC75709, MGC89066, DKFZp468F1016, MGC128356, CYCSA, MGC93634, Cyt-c1, CytC-1, CytC1, Cytc, Cytc-d, DC3, bln, bs30f07.y1, cyt c-d, cyt-c-d, cyt.c, cytochrome c, ms(2)04445, pDMc02, DmelCG13263, CG13263, Cyt-c2, CytC-2, Cytc-p, DC4, DMc01, cyt c-p, cyt-c-p, l(2)SH1590, pDMc01, DmelCG17903, CG17903

Anwendungen

Applikationshinweise: SDS-PAGE & Western Blotting: 1) Wash the cells 3 times with PBS and suspend with 10 volumes of cold Lysis buffer (50 mM Tris-HCl, pH 7.2, 250 mM NaCl, 0.1% NP-40, 2 mM EDTA, 10% glycerol) containing appropriate protease inhibitors. Incubate it at 4 oC with rotating for 30 minutes, then sonicate briefly (up to 10 seconds). 2) Centrifuge the tube at 12,000 x g for 10 minutes at 4 oC and transfer the supernatant to another tube. Measure the protein concentration of the supernatant and add the cold Lysis buffer to make an 8 mg/mL solution. 3) Mix the sample with equal volume of Laemmli’s sample buffer. 4) Boil the samples for 2 minutes and centrifuge. Load 10 μL of the sample per lane in a 1 mm thick SDS-polyacrylamide gel for electrophoresis. 5) Blot the protein to a polyvinylidene difluoride (PVDF)membrane at 1 mA/cm2 for 1 hour in a semi-dry transfer system (Transfer Buffer: 25 mM Tris, 190 mM glycine, 20% MeOH). See the manufacture's manual for specific transfer procedure. 6) To reduce nonspecific binding, soak the membrane in 5% skimmed milk (in PBS, pH 7.2) for 1 hour at room temperature, or overnight at 4 oC. 7) Incubate the membrane with primary antibody diluted with PBS, pH 7.2 containing 1% skimmed milk as suggested in the APPLICATIONS for 1 hour at room temperature. (The optimal antibody concentration will depend on the experimental conditions.) 8) Wash the membrane with PBS (5 minutes x 6 times). 9) Incubate the membrane with the 1:5,000 HRP-conjugated anti-chicken IgY (MBL; code no. PM010-7) diluted with 1% skimmed milk (in PBS, pH 7.2) for 1 hour at room temperature. 10) Wash the membrane with PBS (5 minutes x 6 times). 11) 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 a paper towel, and seal it in plastic wrap. 12) Expose to X-ray film in a dark room for 5 minutes. Develop the film as usual. The conditions for exposure and development may vary. Positive controls for Western blotting; HeLa, NIH/3T3, Rat-1

Buffer: 1 mL aliquot of concentrate antibody from the supernatant with preservative (0.1% sodium azide)

Lagerung: This antibody solution is stable for 3 years from the date of manufacture when stored at 4°C.

Forschungsgebiet: Metabolismus

Beschränkungen: Nur für Forschungszwecke einsetzbar

Bilder

Western blot analysis of cytochrome c expression in HeLa cells (1), Raji cells (2), NIH/3T3 cells (3), CHO cells (4) and Rat-1 cells (5) using ABIN130465