Cytochrome C Human ELISA Kit

Protocol tips

Upstream tips	Protocol tips	Downstream tips
-Allow the microtiter plates to equilibrate to room temperature before opening the foil bags.	-Run a standard curve with each assay. -Drain residual wash liquid from the wells by efficient aspiration or by decantation followed by tapping the plate forcefully on absorbent paper. Never insert absorbent paper directly into the wells. Do not cover plate with foil.	-Once the desired number of strips are removed, immediately reseal the bag and store the plate at 2 to 8°C to maintain plate integrity. -Store both the concentrate and the Working Wash Buffer in the refrigerator. Use the diluted buffer within 14 days.

Upstream tips
-Allow the microtiter plates to equilibrate to room temperature before opening the foil bags.
Protocol tips
-Run a standard curve with each assay. -Drain residual wash liquid from the wells by efficient aspiration or by decantation followed by tapping the plate forcefully on absorbent paper. Never insert absorbent paper directly into the wells. Do not cover plate with foil.
Downstream tips
-Once the desired number of strips are removed, immediately reseal the bag and store the plate at 2 to 8°C to maintain plate integrity. -Store both the concentrate and the Working Wash Buffer in the refrigerator. Use the diluted buffer within 14 days.

Publication protocol

Total protein concentration was determined in triplicate using the Pierce BCA Protein Assay Kit (Thermo Scientific, Waltham, MA) with albumin as a standard. Skeletal muscle citrate synthase (CS), cytochrome C (CytC), and succinate dehydrogenase (SDH) content were determined by an enzyme-linked immunosorbent assay (ELISA; CytC: Thermo Fisher Scientific, Waltham, MA, CAT# KHO1051; CS: GBiosciences, St. Louis, MO, CAT# IT3053; SDH: GBiosciences, St. Louis, MO, CAT# IT4343) in duplicate using a microplate reader (SpectraMax 384 Plus and SoftMax Pro Software, Molecular Devices, San Jose, CA). Intra-assay coefficients of variability (CV) for CS, CytC, and SDH were 4.62, 5.20, and 11.98%, respectively. Mitochondrial protein content was expressed relative to the total protein concentration (ng/mg).

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Manufacturer protocol

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