Cytochrome c Profiling ELISA Kit (ab110172)

Protocol tips

Upstream tips	Protocol tips	Downstream tips
-Equilibrate all reagents to room temperature (18-25°C) prior to use.	-Avoid foaming or bubbles when mixing or reconstituting components. -Ensure plates are properly sealed or covered during incubation steps. -Complete removal of all solutions and buffers during wash steps is necessary to minimize background.	-Unused well strips should be returned to the plate packet and stored at 4°C.

Upstream tips
-Equilibrate all reagents to room temperature (18-25°C) prior to use.
Protocol tips
-Avoid foaming or bubbles when mixing or reconstituting components. -Ensure plates are properly sealed or covered during incubation steps. -Complete removal of all solutions and buffers during wash steps is necessary to minimize background.
Downstream tips
-Unused well strips should be returned to the plate packet and stored at 4°C.

Publication protocol

Cells were cultured at 2×104 cells/well onto 24-well plates. Cell extracts were prepared using cell lysis buffer and centrifuged at 10,000 × g for 15 min at 4°C. The supernatants (200 µl) were detected using a cytochrome c ELISA kit (cat. no. ab110172; Abcam, Cambridge, MA, USA). The assay was performed according to the manufacturer's protocol. Cytochrome c from the conditioned medium was immunocaptured within the wells, and the concentration was determined by adding a cytochrome c-specific antibody conjugated to horseradish peroxidase. Following addition of the provided colorless substrate, the peroxidase converted the substrate to a blue end-product. This reaction occurred in a time-dependent manner, which was proportional to the amount of protein captured in the wells. The rate of blue color development was detected at 600 nm. The change in absorbance was expressed as change in milliOD/min.

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Manufacturer protocol

Download the product protocol from Abcam for Cytochrome c Profiling ELISA Kit (ab110172) below.

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