Preparation of the Samples for Cytochrome c ELISA

Materials

• Quantikine® Rat/Mouse Cytochrome c ELISA Kit (R&D Systems Catalog # MCTC0)
• 5% Triton® X-100

Methods

1. Thaw the samples immediately prior to running the ELISA.
2. Add 7.5 µL of 5% Triton® X-100 to the total Cytochrome c control sample (75 µL) that was frozen immediately after incubation at 30° C.
3. Add 5.0 µL of 5% Triton X-100 to the remaining 50 µL samples and controls. Vortex the samples.
4. Dilute all samples and controls 100-fold with Cytochrome c ELISA diluent, RD5-18. Vortex the samples.
5. Assay 50 µL aliquots of the 100-fold diluted samples in triplicate in the 96-well plate included in the Rat/Mouse Cytochrome c ELISA Kit (Catalog # MCTC0) according to the kit instructions.

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Triton is a registered trademark of Union Carbide Corp.