Sony Biotechnology Inc. Sony Biotechnology Inc.cart
About Us Products What's New Library News Calendar Contact Us Place Order
Safety Data Sheet (SDS) Purified Anti-Cytochrome C Antibody     Product Data Sheet (PDF)    
Purified Anti-Cytochrome C Antibody
3661505 25 µg $75.00       
3661510 100 µg $165.00       
Clone: 6H2.B4
Isotype: Mouse IgG1, κ
Reactivity: Mouse, Rat, Human
Immunogen: Rat cyt c-OVA
Formulation: This antibody is provided in phosphate-buffered solution, pH 7.2, containing 0.09% sodium azide. Final antibody concentration is 0.5 mg/ml.
Preparation: The antibody was purified by affinity chromatography.
Concentration: 0.5 mg/ml
Storage & Handling: Upon receipt, store between 2°C and 8°C.
Application:

IP - Quality tested
IF - Validated
ICFC, ICC - Reported in the literature

Application Notes:

Additional reported applications (for the relevant formats) include: intracellular flow cytometry5, immunofluorescence microscopy3,5, immunoprecipitation4, and immunocytochemistry5.

Recommended Usage:

Each lot of this antibody is quality control tested by immunoprecipitation. This antibody can be used at 2-4 µg /1 x107 cell equivalents for immunoprecipitation. For flow cytometric staining, the suggested use of this reagent is ≤0.5 µg per 106 cells in 100 µl volume.
It is recommended that the reagent be titrated for optimal performance for each application.

Application References:

1. Goshorn SC, et al. 1991. J. Biol. Chem. 266:2134.
2. Jemmerson R, et al. 1991. Eur. J. Immunol. 21:143.
3. Chandra D, et al. 2002. J. Biol. Chem. 277:50842. (IF)
4. Semenkova L, et al. 2003. Eur. J. Biochem. 270:4388. (IP)
5. Shih S-F, et al. 2001. J. Biol. Chem. 276:21870. (ICFC ICC IF)
6. She P, et al. 2011. Am J. Physiol Endcorinol Metab.301:E49. PubMed
7. McGuire, KA., et al. 2011. J. Virol 85:10806. PubMed

Immunoprecipitation/Western Blot analysis using purified

Immunoprecipitation/Western Blot analysis using purified anti-Cytochrome c antibody (6H2.B4). 800 µg of HeLa cell lysates was tested at protein concentration of 1µg/µl for each sample. Lane 1 was immunoprecipitated with control antibody and lane 2 was immunoprecipitated with anti-Cytochrome c Antibody (6H2.B4). Western blot analysis was performed using anti-Cytochrome c Antibody (7H8.2C12).

HeLa cells were fixed with

HeLa cells were fixed with 1% paraformaldehyde (PFA) for 10 minutes, permeabilized with 0.5% Triton X-100 for 10 minutes, and blocked with 5% FBS for 30 minutes. Then the cells were intracellularly stained with 5 µg/ml of purified cytochrome c (clone 6H2.B4) in blocking buffer overnight at 4°C, and followed by DyLight™ 594 anti-mouse IgG (red) staining for 2 hours at 4°C. Nuclei were counterstained with DAPI and are shown in blue. The image was captured with 40X objective.



Description:

Cytochrome c is a 15 kD protein found in the mitochondrial intermembrane space with a heme-binding domain. Cytochrome c is a component of the electron transport chain; the heme group transfers electrons from cytochrome b-c1 complex to cytochrome oxidase complex. Cytochrome c initiates apoptosis by release to cytoplasm and binding Apaf-1 which activates procaspase 9. Cytochrome c interacts with the cytochrome b-c1 complex, cytochrome oxidase complex, heme, Apaf-1, and Caspase 9 proteins. The 6H2.B4 monoclonal antibody recognizes human, mouse, and rat cytochrome-c and has been shown to be useful for intracellular flow cytometric staining, Western blotting, immunoprecipitation, and immunofluorescence staining.

Other Names: Cyt c
Structure: Heme binding domain; 15 kD
Distribution: Mitochondrial intermembrane space
Function: Component of electron transport chain; heme group transfers electrons from cytochrome b-c1 complex to cytochrome oxidase complex. Initiates apoptosis by release to cytoplasm and binding Apaf-1 which activates procaspase 9
Interaction: Cytochrome b-c1 complex, cytochrome oxidase complex, heme, Apaf-1, Casp9
Antigen References:

1. Liu X, et al. 1996. Cell. 86:147.
2. Li P, et al. 1997. Cell. 91:479.
3. Zhang Z, et al. 2003. Gene 312:61.
4. Ferguson H, et al. 2003. J. Biol. Chem. 278:45793.