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Our lab want to purchase caspase 1 (ab39901) and caspase 3 (ab52101) for
my project. We have one question regarding these products. We want to know
if we are going to cut our protein (40ug) with caspase 1 or 3, how many ul of
caspase 1 or 3 we need to add into the buffer in order to have a useful reaction?
Another question is I checked your website but I only found 25 units in caspase 1
and 100 units in caspase 3. Can I have the information about how many ul or ml
you have in one tube of caspase 1 or 3?
Thank you for all the information.
Asked on Feb 21 2012
Thank you for contacting Abcam.
For ab39901, Caspase 1, we recommend using 1 unit/assay for analyzing caspase activity and this protein has a biological activity : >5000 units/mg and it is supplied lyophilized and it should be reconstituted to 1 unit per µl in PBS containing 15% glycerol (and as it supplied as 25 units you would end up with 25ul of solution).
For ab52101, Caspase 3, one unit of the recombinant caspase-3 is the enzyme activity that cleaves 1 nmol of the caspase substrate DEVD-pNA (pNA: pnitroanaline) per hour at 37oC in a reaction solution containing 50 mM Hepes, pH 7.2, 50 mM NaCl, 0.1% Chaps, 10 mM EDTA, 5% Glycerol, and 10 mM DTT. You are supplied with 100 units and again it is lyophilized on arrival an you should reconstitute to 1 unit per µl in PBS containing 15% glycerol (therefore you would have 100ul of solution).
The above are recommended starting dilutions, however the optimal dilutions/concentrations should be determined by the end user.
Please let me know if there is anything else I can help you with.
Answered on Feb 21 2012