< 0.5% cross-reactivity observed with available related molecules.< 50% cross-species reactivity observed with species tested.
Interference observed with other substances.
The Quantikine Human Cytochrome c Immunoassay is a 4.5 hour solid-phase ELISA designed to measure human cytochrome c in cell lysates.
Intra-Assay Precision (Precision within an assay) Three samples of known concentration were tested on one plate to assess intra-assay precision.
Inter-Assay Precision (Precision between assays) Three samples of known concentration were tested in separate assays to assess inter-assay precision.
Cell Lysates, Subcellular Fractions
To assess the linearity of the assay, six cell lysates containing high concentrations of Cytochrome c were diluted with Calibrator Diluent to produce samples with values within the dynamic range of the assay.
Preparation and Storage
Store the unopened product at 2 - 8 °C. Do not use past expiration date.
Background: Cytochrome c
Cytochrome c is a small, highly conserved, heme-containing protein involved in mitochondrial electron transport. Cytochrome c is released from the mitochondria in response to pro-apoptotic stimuli and serves as a cofactor in initiating the activation of the APAF-1/Caspase-9 complex. The effect of specific proteins and/or small molecules on the initiation of the intrinsic apoptotic pathway can be evaluated in an in vitro system of isolated mitochondria by monitoring the release of cytochrome c.
Refer to the product for complete assay procedure.
Bring all reagents and samples to room temperature before use. It is recommended that all samples, standards, and controls be assayed in duplicate.
Prepare all reagents, standard dilutions, and samples as directed in the product insert.
Remove excess microplate strips from the plate frame, return them to the foil pouch containing the desiccant pack, and reseal.
100 µL Calibrator Diluent
Add 100 µL of Calibrator Diluent to each well.
100 µL Standard, Control, or Sample
Add 100 µL of Standard, control, or sample to each well. Cover with a plate sealer, and incubate at room temperature for 2 hours.
Aspirate each well and wash, repeating the process 3 times for a total of 4 washes.
200 µL Conjugate
Add 200 µL of Conjugate to each well. Cover with a new plate sealer, and incubate at room temperature for 2 hours.
Aspirate and wash 4 times.
200 µL Substrate Solution
Add 200 µL Substrate Solution to each well. Incubate at room temperature for 30 minutes. PROTECT FROM LIGHT.
50 µL Stop Solution
Add 50 µL of Stop Solution to each well. Read at 450 nm within 30 minutes. Set wavelength correction to 540 nm or 570 nm.
R&D Systems personnel manually curate a database that contains references using R&D Systems products.
The data collected includes not only links to publications in PubMed,
but also provides information about sample types, species, and experimental conditions.