Amresco
VisiGlo Prime
1B1581-KIT-SAMPLE
VisiGlo Prime was assessed as a chemiluminescence reagent for Western blotting. We sought to compare this reagent to other HRP substrates, and determine their signal intensity and background.
Western blotting
Western blot
20 ug of cytoplasmic lysates from human B lymphoblastoid cell lines were electrophoresed on a 4-15% Bio-Rad TGX gel for 35 mins and transferred to a PVDF membrane using standard protocols. Blots were probed with rabbit anti-human GAPDH antisera (Santa Cruz Biochemicals) at a 1:500 dilution for 30 mins followed by goat-anti-rabbit HRP secondary antibodies at 1:1000 (Santa Cruz) using SWIFT™ Western Diluent (G-Biosciences) according to the manufacturer’s protocol. Following 3 5 min washes in TBST, a 1:1 mixture of VisiGlo Prime reagent was added to the blot and exposed for 2-10 seconds in a Bio-Rad Chemi-Doc imager. A 4 second exposure is shown in the figure.
Small amounts go a long way. The blot shown in the Figure has only 500 ul of the reagent, and we probably could have used less.
We obtained a publication quality image in two to four seconds. Background levels were higher than a competitor's substrate, but this can probably be avoided by using more washing steps after the secondary antibody is used.
I suspect that VisiGlo may be best used with weak or low quantities of antibodies, maximizing bang for the buck.
Rapid results, easy to use
Cost! List price is $245 for 100ml; Bio-Rad's Clarity substrate has 200 ml for $130
Excellent signal and rapid results.