We tried to quantify Caspase 3 activity in mouse mammary gland during mammary involution after lactation. We tested different protein amounts fron 50 to 200 microgrames (as indicated in protocol booklet), but the product did not work in any case. We wanted to compare Caspase 3 activity in three samples at different timepoints of involution, and we were not able to observe differences (A), which really exist, as we confirmed by western blot of the same samples (B). We checked the kit was not faulty testig in HC11 mammary cell line, inducing apoptosis with cisplatin 15 ug/ml for 24h (C). The technical service told us it was a problem with our samples and the did not offer a way to solve our problem. Then, we found a fluorimetric assay ab39383, which is exactly the same but with a fluorometric substrate instead of a colorimetric one, which is more sensitive as abcam scientific support says in the questions and answer section of the product wall in their website. We aquired the fluorometric substrate and repeated the assay with the same samples. This time, the product worked perfectly (D). So, for mouse mammary gland tissue samples, I would not recommend this product. It seems it has not enough sensitivity. I would try the fluorimetric assay.
Mr. Adrian Blanco
Submitted Jul 17 2014
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