Mouse/Rat Cytochrome C ELISA Kit (ab210575)
- Datasheet
- References
- Protocols
Overview
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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.
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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.
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Tested applications
Suitable for: Sandwich ELISAmore details -
Platform
Microplate (12 x 8 well strips)
Properties
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Storage instructions
Store at +4°C. Please refer to protocols. -
Components 1 x 96 tests 10X Mouse/Rat Cytochrome C Capture Antibody 1 x 600µl 10X Mouse/Rat Cytochrome C Detector Antibody 1 x 600µl 10X Wash Buffer PT (ab206977) 1 x 20ml 50X Cell Extraction Enhancer Solution (ab193971) 1 x 1ml 5X Cell Extraction Buffer PTR (ab193970) 1 x 10ml Antibody Diluent 5BR 1 x 6ml Denaturant 1 x 500µl Mouse Cytochrome C Lyophilized Recombinant Protein 2 vials Plate Seals 1 unit Sample Diluent NS (ab193972) 1 x 12ml SimpleStep Pre-Coated 96-Well Microplate (ab206978) 1 unit Stop Solution 1 x 12ml TMB Development Solution 1 x 12ml -
Research areas
- Metabolism
- Pathways and Processes
- Metabolic signaling pathways
- Lipid and lipoprotein metabolism
- Lipases
- Metabolism
- Pathways and Processes
- Metabolic signaling pathways
- Energy transfer pathways
- Energy Metabolism
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Function
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.
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 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. -
Involvement in disease
Defects in CYCS are the cause of thrombocytopenia type 4 (THC4) [MIM:612004]; also known as autosomal dominant thrombocytopenia type 4. Thrombocytopenia is the presence of relatively few platelets in blood. THC4 is a non-syndromic form of thrombocytopenia. Clinical manifestations of thrombocytopenia are absent or mild. THC4 may be caused by dysregulated platelet formation. -
Sequence similarities
Belongs to the cytochrome c family. -
Post-translational
modificationsBinds 1 heme group per subunit. -
Cellular localization
Mitochondrion matrix. - Information by UniProt
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Alternative names
- CYC
- CYC_HUMAN
- CYCS
see all -
Database links
- Entrez Gene: 13063 Mouse
- SwissProt: P62897 Mouse
- Unigene: 35389 Mouse
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
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Other - Mouse/Rat Cytochrome C ELISA Kit (ab210575)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.
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Mouse/Rat Cytochrome C standard curve comparison data.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.
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Example of mouse/rat Cytochrome C standard curve.Background-subtracted data values (mean +/- SD) are graphed.
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Interpolated concentrations of native Cytochrome C in mouse heart tissue extract samples based on 15 µg/mL extract load and in rat heart tissue extract samples based on 7 µg/mL extract load.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.
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Interpolated concentrations of native Cytochrome C in mouse NIH/3T3 cell extract samples based on 65 µg/mL extract load, and mouse C2C12 and rat C6 cell extract samples based on 100 µg/mL extract.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.
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Interpolated concentrations of native Cytochrome C in mouse and rat extract samples.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.
