Describe the experiment that the product used for. If an antibody, what application and protocol was used?
EpiQuik Invivo Universal protein Sumoylation Assay Kit, Cat No. P-8001
How friendly was the user guide? (1 – worst, 5 – best)
User guide is best, If treated and untreated sample can be define better that would be great
How professional was the appearance and presentation of the product? (1 – worst, 5 – best)
appearance and presentation of the product is best
How would you rate the overall product? (1 – Worst, 5 – best)
Pros: Easy and faster to show in Vivo Sumoylation, Required small amount of protein
Cons: there was no cons of the product
Sensitive Sumo-ELISA/Sandwich-ELISA assays validated that both extrinsic and intrinsic Prdx6 were Sumoylated, and showed that a fraction of endogeneous Prdx6 was present in Sumoylated form. (A) hLECs were transfected with pEGFP-vector or pEGFP-vector plus pEGFP-Sumo1 or pGFP-Prdx6 plus pEGFP-Sumo1. After 48h total cell lysates were prepared and submitted to Sandwich/Sumo1-ELISA assays to check the total Prdx6 protein and Sumoylated Prdx6 protein. Sumoylated Prdx6 protein was subtracted from total Prdx6 protein, presenting as Prdx6 unSumoylated (gray bars) and Sumoylated (black bars) forms. The data represent mean ± SD from three independent experiments (*p<0.001). (B) Prdx6+/+ LECs were transfected with pEGFP-vector plus pEGFP-Sumo1 or pGFP-Prdx6 plus pEGFP-Sumo1. Total cell lysates were prepared and used to perform Sandwich/Sumo1-ELISA. Sumoylated Prdx6 protein was subtracted from total Prdx6 protein, presenting as Prdx6 unSumoylated (gray bars) and Sumoylated (black bars) forms. The data represent the mean ± SD of three independent experiments (*p<0.001 vs control).
Overall kit is ok but company providing very less Sumoylation buffer. As now its known that Sumoylation occurs in nucleus as well as cytosol so total lysate or nuclear protein can be used. As many labs use own lysis buffer (RIPA) to isolate protein. Sometime protein concentration is low (eg 0.5ug/ul or 1ug/ul) so we have to increase the volume of protein so of course have to increase SUMO assay buffer. All buffer volume is ok but Sumoylation assay buffer volume is too low. Company can increase the volume of Sumoylation assay buffer or can provide the method.