Caspase 3 assay - on purified proteins (Sep/23/2008 )
I want to seek the cleave of a purified protein with caspase-3. I have purchased recombinant caspase from Sigma (55uL of 0.25mg/ml; activity:5.670U/mg (sigma 1 unit definition= cuts 1ug of caspase 3 peptide subs).
Purified protein is histagged expressed in E.coli.
So I was what should be the starting amount of protien and how many units of enzyme.
Is their any specified protocol for such assays, if not then how to proceed, any help.
thnx
Purified protein is histagged expressed in E.coli.
So I was what should be the starting amount of protien and how many units of enzyme.
Is their any specified protocol for such assays, if not then how to proceed, any help.
thnx
you mean you purchased the protein without even seeking info before buying it? then where did you hear about this protein and method in first place?
please if you dont understand me or dont know answer then you should not write anything. I simply asking the amount of starting protein detectable on western after cleavage, and was asking if somebody has done here assay, this is very common assay for the people who work to look for new substrates of caspases but I never worked with both pure proteins, so was the question addressed.
well, i think you need to work on your English before posting here, otherwise nobody can answer your question. we load 30 ug of protein on each well usually.
well, i think you need to work on your English before posting here, otherwise nobody can answer your question. we load 30 ug of protein on each well usually.
thanks for reply and you are right my English is terrible, trying hard to improve.