Review text: Dilutions of PDE12 (1 to 10ml) were incubated for 2h with 20µl of PDE12 assay substrate, an analogue of 2-5A dual labelled with Fam and Dabcyl. Active PDE12 is expected to cleave 2-5A molecules releasing the quencher from the Fluorophore, allowing it to fluoresce which is detected by the Flo-Star Optima (Ex 485nm and Em 520nm). Delta fluorescence (T120min-Tzero min) is used to calculate PDE12 % activity using cell lysate as positive control for PDE12.
Unfortunately there was no detectable fluorescence (Tzero and T120 were almost the same) observed after incubating PDE12 (ab107695) for 2h suggesting the protein under the storage conditions provided was not active.
Sample: Human Purified protein
Primary antibody (in addition to 'PDE12 protein')
Primary antibody: None used
Secondary antibody: None used
Abcam response: Accordingly to these results, this protein preparation has not conserved its functional activity. We thank the researcher to have participated in our testing offer. Unfortunately we have to conclude that this protein in ists current formulation seems not suitable for functional tests.
Dr. fouzia sadiq
Submitted Sep 16 2011