Overview

  • Product name

    Mouse/Rat Cytochrome C ELISA Kit
    See all Cytochrome C kits
  • Detection method

    Colorimetric
  • Precision

    Intra-assay
    Sample n Mean SD CV%
    NIH3T3 ext 5 2.7%
    Inter-assay
    Sample n Mean SD CV%
    NIH3T3 ext 3 5.3%
  • Sample type

    Cell culture extracts, Tissue Extracts, Cell culture media
  • Assay type

    Sandwich (quantitative)
  • Sensitivity

    44 pg/ml
  • Range

    487.4 pg/ml - 14000 pg/ml
  • Recovery

    Sample specific recovery
    Sample type Average % Range
    Tissue Extracts 108 106% - 112%
    Cell culture media 111 106% - 116%

  • Assay time

    1h 30m
  • Assay duration

    One step assay
  • Species reactivity

    Reacts with: Mouse
  • Product overview

    Abcam’s Cytochrome C in vitro SimpleStep ELISA® (Enzyme-Linked Immunosorbent Assay) kit is designed for the quantitative measurement of Cytochrome C protein in mouse and rat cell and tissue extracts.


    The SimpleStep ELISA® employs an affinity tag labeled capture antibody and a reporter conjugated detector antibody which immunocapture the sample analyte in solution. This entire complex (capture antibody/analyte/detector antibody) is in turn immobilized via immunoaffinity of an anti-tag antibody coating the well. To perform the assay, samples or standards are added to the wells, followed by the antibody mix. After incubation, the wells are washed to remove unbound material. TMB substrate is added and during incubation is catalyzed by HRP, generating blue coloration. This reaction is then stopped by addition of Stop Solution completing any color change from blue to yellow. Signal is generated proportionally to the amount of bound analyte and the intensity is measured at 450 nm. Optionally, instead of the endpoint reading, development of TMB can be recorded kinetically at 600 nm.

  • Notes

    Cytochrome C is 11 kDa mitochondrial intermembrane space electron carrier protein. The oxidized form of the cytochrome C heme group can accept an electron from the heme group of the cytochrome c1 subunit of cytochrome reductase. Cytochrome C then transfers this electron to the cytochrome oxidase complex, the final protein carrier in the mitochondrial electron-transport chain.

    Cytochrome C also plays a role in apoptosis. Suppression of the anti-apoptotic members or activation of the pro-apoptotic members of the Bcl-2 family leads to altered mitochondrial outer membrane permeability resulting in release of cytochrome C into the cytosol. Binding of cytochrome C to Apaf-1 triggers the activation of caspase-9, which then accelerates apoptosis by activating other caspases.

  • Tested applications

    Suitable for: Sandwich ELISAmore details
  • Platform

    Microplate (12 x 8 well strips)

Properties

Applications

Our Abpromise guarantee covers the use of ab210575 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Application Abreviews Notes
Sandwich ELISA Use at an assay dependent concentration.

Images

  • SimpleStep ELISA technology allows the formation of the antibody-antigen complex in one single step, reducing assay time to 90 minutes. Add samples or standards and antibody mix to wells all at once, incubate, wash, and add your final substrate. See protocol for a detailed step-by-step guide.

     

  • Standard curve comparison between mouse/rat Cytochrome C SimpleStep ELISA® kit and traditional ELISA kit from leading competitor. SimpleStep ELISA kit shows a 10-fold increase in sensitivity.

  • Background-subtracted data values (mean +/- SD) are graphed.

  • The concentrations of Cytochrome C were measured in duplicate and interpolated from the Cytochrome C standard curve and corrected for sample dilution. The interpolated dilution factor corrected values are plotted (mean +/- SD, n=2). The mean Cytochrome C concentration was determined to be 8,822 pg/mL in mouse heart extract and 4,109 pg/mL in rat heart extract samples.

  • The concentrations of Cytochrome C were measured in duplicate and interpolated from the Cytochrome C standard curve and corrected for sample dilution. The interpolated dilution factor corrected values are plotted (mean +/- SD, n=2). The mean Cytochrome C concentration was determined to be 10,516 pg/mL in NIH/3T3 extract, 11,096 pg/mL in C2C12 extract, and 13,297 pg/mL in C6 extract.

  • The concentrations of Cytochrome C were measured in three different dilutions in duplicate and interpolated from the Cytochrome C standard curve and corrected for sample dilution. The interpolated dilution factor corrected values are plotted in ng of Cytochrome C per mg of extract (mean +/-SD, n=3). Cytochrome C concentration was determined to be 601 ng/mg in mouse heart, 622 ng/mg in rat heart, 166 ng/mg in mouse NIH/3T3 cell, 105 ng/mg in mouse C2C12 cell and 136 ng/mg in rat C6 cell extract samples.

Protocols

References

ab210575 has not yet been referenced specifically in any publications.

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