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

Measurement of liver and kidney caspase-3 activity was performed using the Caspase-3 Colorimetric Assay Kit (Abcam, ab39401, Cambridge, UK) following the manufacturer's instructions. Livers and kidneys were cut into small pieces, washed three times with supplied cell lysis buffer and homogenized with a tissue grinder. Liver and kidney homogenates were centrifuged for 10 min at 12 000 r.p.m. and supernatants were used for colorimetric determination at 405 nm. The OD values were normalized to the protein levels present in the tissues. The homogenates were also used to isolate cytosolic fraction with Standard Cell Fractionation Kit (Abcam, ab109719) and cytosolic CytC levels were determined by Cytochrome c Profiling ELISA Kit (Abcam, ab110172) and normalized to protein levels present in the tissues.

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Manufacturer protocol

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

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