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.
Human Cytochrome c Quantikine ELISA Kit
R&D Systems | Catalog # DCTC0
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Key Product Details
Assay Length
4.5 hours
Sample Type & Volume Required Per Well
Cell Lysates (20 µL), Subcellular Fractions (100 µL)
Sensitivity
0.31 ng/mL
Assay Range
0.625-20 ng/mL (Cell Lysates, Subcellular Fractions)
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Product Summary for Human Cytochrome c Quantikine ELISA Kit
The Quantikine Human Cytochrome c Immunoassay is a 4.5 hour solid-phase ELISA designed to measure human cytochrome c in cell lysates.
Product Specifications
Assay Type
Solid Phase Sandwich ELISA
Format
96-well strip plate
Measurement
Quantitative ELISA
Detection Method
Colorimetric - 450nm (TMB)
Conjugate
HRP
Species
Human
Specificity
Natural human Cytochrome c
Cross-reactivity
< 0.5% cross-reactivity observed with available related molecules. < 50% cross-species reactivity observed with species tested.
Interference
Interference observed with other substances.
Precision
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
| Intra-Assay Precision | Inter-Assay Precision | |||||
|---|---|---|---|---|---|---|
| Sample | 1 | 2 | 3 | 1 | 2 | 3 |
| n | 20 | 20 | 20 | 40 | 40 | 40 |
| Mean (pg/mL) | 3.64 | 7.31 | 10.7 | 3.54 | 6.74 | 10.2 |
| Standard Deviation | 0.15 | 0.30 | 0.42 | 0.18 | 0.34 | 0.51 |
| CV% | 4.1 | 4.1 | 3.9 | 5.1 | 5.0 | 5.0 |
Linearity
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.
Scientific Data Images for Human Cytochrome c Quantikine ELISA Kit
Human Cytochrome c ELISA Standard Curve
Preparation and Storage
Shipping
The product is shipped at ambient temperature. Upon receipt, store it immediately at the temperature recommended below.
Stability & Storage
Store the unopened product at 2 - 8 °C. Do not use past expiration date.
Background: Cytochrome c
Additional Cytochrome c Products
Product Documents for Human Cytochrome c Quantikine ELISA Kit
Certificate of Analysis
To download a Certificate of Analysis, please enter a lot or batch number in the search box below.
Note: Certificate of Analysis not available for kit components.
Product Specific Notices for Human Cytochrome c Quantikine ELISA Kit
For research use only
⚠ WARNING: This product can expose you to chemicals including N,N-Dimethylforamide, which is known to the State of California to cause cancer. For more information, go to www.P65Warnings.ca.gov.Related Research Areas
Citations for Human Cytochrome c Quantikine ELISA Kit
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Protocols
View specific protocols for Human Cytochrome c Quantikine ELISA Kit (DCTC0):
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.
- Add 100 µL of Calibrator Diluent to each well.
- 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.
- 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.
- Add 200 µL Substrate Solution to each well. Incubate at room temperature for 30 minutes. PROTECT FROM LIGHT.
- 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.
100 µL Calibrator Diluent
100 µL Standard, Control, or Sample
200 µL Conjugate
200 µL Substrate Solution
50 µL Stop Solution
Find general support by application which include: protocols, troubleshooting, illustrated assays, videos and webinars.
- ELISA Sample Preparation & Collection Guide
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- View all Protocols, Troubleshooting, Illustrated assays and Webinars
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Associated Pathways
MAPK Signaling: Oxidative Stress Pathway
