Purified anti-Cytochrome c Antibody

Pricing & Availability
Clone
7H8.2C12 (See other available formats)
Other Names
Cyt c
Isotype
Mouse IgG2a, κ
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Product Citations
publications
1_7H8dot2C12_PURE_CytochromeC_WB_Antibody_060118
Total lysates (15 µg protein) from HeLa (Human), Raw264.7 (Mouse), UMR106 (Rat) and CHO (Hamster) were resolved by electrophoresis (4-20% Tris-glycine gel), transferred to nitrocellulose, and probed with 1:1000 purified anti-Cytochrome C antibody, clone 7H8.2C12. Proteins were visualized using chemiluminescence detection by incubation with HRP Goat anti-Mouse secondary antibody (Cat. No. 405306, 1:3000 dilution). Non specific band was marked with “*”. Direct-Blot™ HRP anti-β-actin was used as a loading control (Cat. No. 643807, 1:8000 dilution).
  • 1_7H8dot2C12_PURE_CytochromeC_WB_Antibody_060118
    Total lysates (15 µg protein) from HeLa (Human), Raw264.7 (Mouse), UMR106 (Rat) and CHO (Hamster) were resolved by electrophoresis (4-20% Tris-glycine gel), transferred to nitrocellulose, and probed with 1:1000 purified anti-Cytochrome C antibody, clone 7H8.2C12. Proteins were visualized using chemiluminescence detection by incubation with HRP Goat anti-Mouse secondary antibody (Cat. No. 405306, 1:3000 dilution). Non specific band was marked with “*”. Direct-Blot™ HRP anti-β-actin was used as a loading control (Cat. No. 643807, 1:8000 dilution).
  • 2_7H8dot2C12_PURE_CytochromeC_Antibody_IF_082917
    Hela cells were pre-incubated with 400nM MitoSpy™ Green FM at 37°C for 30 minutes and then fixed with 4% paraformaldehyde (PFA) for 15 minutes, permeabilized with 0.5% Triton X-100 for 3 minutes, and blocked with 5% FBS for 60 minutes. The cells were intracellularly stained with either (A) 3 µg/ml anti-mouse IgG1, κ, or (B) anti-cytochrome C antibody (clone 7H8.2C12), and incubated overnight at 4°C followed by 27micro;g/ml (1:250) Alexa Fluor® 594 (Red) goat anti-mouse IgG for one hour at room temperature. Nuclei were counterstained with DAPI. The image was captured with a 60X objective. Exposure time for (A) is 1/30, and (B) is 1/60.
  • 3_7H8dot2C12_PURE_CytochromeC_Antibody_IF_2_121217
    HeLa cells were treated with 400 nM MitoSpy™ Green FM (Green, Cat. No. 424805) for 30 minutes, fixed with 4% paraformaldehyde (PFA) for fifteen minutes, permeabilized with 0.5% Triton X-100 for three minutes, and blocked with 5% FBS for 60 minutes. Then the cells were intracellularly stained with purified anti-Cytochrome c antibody (clone 7H8.2C12) overnight at 4°C followed by Alexa Fluor® 594 (Red) goat anti-mouse IgG for one hour at room temperature (Cat. No. 405326, 1:250 dilution, 2 µg/ml). Nuclei were counterstained with DAPI (blue, Cat. No. 422801). The image was captured with a 60X objective. The image was captured with a 60X objective using KEYENCE BZ-X700 fluorescence microscope. Exposure time (in seconds) is 1/20.
  • 7H8dot2C12_PURE_CytochromeC_Antibody_IF_3_121217
    HeLa cells were treated with 400 nM MitoSpy™ Red CMXRos (Red, Cat. No. 424801) for 30 minutes, fixed with 4% paraformaldehyde (PFA) for fifteen minutes, permeabilized with 0.5% Triton X-100 for three minutes, and blocked with 5% FBS for 60 minutes. Then the cells were intracellularly stained with purified anti-Cytochrome c antibody (clone 7H8.2C12) overnight at 4°C followed by Alexa Fluor® 488 (Green) goat anti-mouse IgG for one hour at room temperature (Cat. No. 405319, 1:250 dilution, 2 µg/ml). Nuclei were counterstained with DAPI (Blue, Cat. No. 422801). The image was captured with a 60X objective. The image was captured with a 60X objective using KEYENCE BZ-X700 fluorescence microscope. Exposure time (in seconds) is 1/20.
Cat # Size Price Quantity Avail. Save
612503 50 µl (5 Western blots) $90
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612504 200 µl (20 Western blots) $250
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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 7H8.2C12 monoclonal antibody recognizes cytochrome-c from most species and has been shown to be useful for Western blotting.

Product Details
Technical data sheet

Product Details

Reactivity
Most Species
Antibody Type
Monoclonal
Host Species
Mouse
Immunogen
Horse cyt c-OVA
Formulation
This antibody is provided in phosphate-buffered solution, pH 7.2, containing 0.09% sodium azide.
Preparation
The antibody was purified by affinity chromatography.
Concentration
Lot-specific (please contact technical support for concentration and total µg amount, or use our Lookup tool if you have a lot number.)
Storage & Handling
Upon receipt, store undiluted between 2°C and 8°C.
Application

WB - Quality tested
ICC - Validated

Recommended Usage

Each lot of this antibody is quality control tested by Western blotting. Western blotting, suggested working dilution(s): Use 10 µl antibody per 5 ml antibody dilution buffer for each mini-gel. For immunocytochemistry, a concentration range of 1.0 - 3.0 µg/ml (1:150-1:500 dilution) is recommended. It is recommended that the reagent be titrated for optimal performance for each application.

Application References

(PubMed link indicates BioLegend citation)
  1. Jemmerson R, et al. 1991. Eur. J. Immunol. 21:143. (WB)
  2. Semenkova L, et al. 2003. Eur. J. Biochem. 270:4388. (WB)
Product Citations
  1. Duncan H 2010. Infect Immun. 78:3616. PubMed
  2. Asano M, et al. 2017. Sci Rep. 10.1038/s41598-017-11784-y. PubMed
  3. Liu X 2018. Cell. 174:187. PubMed
  4. Kakimoto PA, et al. 2019. PLoS One. 14:e0211733. PubMed
Publication Library
RRID
AB_2090157 (BioLegend Cat. No. 612503)
AB_2292697 (BioLegend Cat. No. 612504)

Antigen Details

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
Biology Area
Apoptosis/Tumor Suppressors/Cell Death, Cell Biology, Mitochondrial Function, Neuroscience, Neuroscience Cell Markers
Molecular Family
Mitochondrial Markers
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.

Gene ID
54205 View all products for this Gene ID
UniProt
View information about Cytochrome c on UniProt.org

Related FAQs

There are no FAQs for this product.
Go To Top Version: 3    Revision Date: 06/01/2018

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This data display is provided for general comparisons between formats.
Your actual data may vary due to variations in samples, target cells, instruments and their settings, staining conditions, and other factors.
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