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Abcam’s Cytochrome C in vitro ELISA (Enzyme-Linked Immunosorbent Assay) kit is designed for the measurement of Cytochrome C in Human, mouse, rat and bovine samples of purified mitochondria, tissue and cell extracts.
Abcam’s Cytochrome C ELISA kit is used to determine the amount of the cytochrome c in a sample. Cytochrome c is immunocaptured within the wells and the amount is determined by adding a cytochrome c specific antibody conjugated with horseradish peroxidase (HRP). This peroxidase changes the substrate from colorless to blue. This reaction takes place in a time dependent manner proportional to the amount of protein captured in the wells. The rate of development of blue color can be followed at 600 nm or the reaction can be stopped, at a user defined time, by the addition of 100 μL of 1.5 N HCl to each well (not supplied) and read at 450 nm.
The permeabilization of the mitochondrial outer membrane and the consequent release of cytochrome c and other apoptogenic proteins from the mitochondrial intermembrane space into cytoplasm is considered hallmark of many apoptotic pathways leading to cell death. Therefore assaying these proteins in mitochondrial and cytoplasmic fractions is a prime interest of many researchers. Most of the current biochemical methods of quantification of cytoplasmic and mitochondrial cytochrome c involve mechanical disruption of cells to obtain mitochondria-enriched and cytosolic fractions. These methods are time-consuming and limited to small number of samples.
This cytochrome c detection assay was developed in conjunction with Abcam’s ab109719 Cell Fractionation Kit - Standard and ab109718 Cell Fractionation Kit - HT, and the results were validated with ab110417 ApoTrack™ Cytochrome c Apoptosis Cocktail and with ab110415 ApoTrack™ Apoptosis Detection Kit for Immunocytochemistry.
|10X Blocking Buffer||1 x 15ml|
|20X Buffer||Tube 1||1 x 15ml|
|20X Detector antibody||Tube A||1 x 1ml|
|20X HRP label||Tube B||1 x 1ml|
|20X Wash Buffer||Tube 3||1 x 15ml|
|96-well pre-coated microplate||1 unit|
|Detergent||1 x 1ml|
|HRP/TMB Development Solution||1 x 20ml|
Our Abpromise guarantee covers the use of ab110172 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|Sandwich ELISA||Use at an assay dependent dilution.|