Advansta
WesternBright Quantum HRP substrate reagent
K-12042-C20
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The replacement of Xray film with digital imaging requires high quality substrates for chemiluminescence detection. We tested WesternBright Quantum substrate in a head-to-head competition with another manufacturer's product.
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, the blot was cut in half. One blot received 500 ul of a competitor’s reagent, while the second received a 1:1 mixture of WesternBright Quantum HRP substrate reagent. The blots were exposed for 2-10 seconds in a Bio-Rad Chemi-Doc imager. A 4 second exposure is shown in the figure.
For the small size of the blot we used, we suspect that a smaller volume of reagent could have been used., perhaps 250-300 ul.
Compared to VisiGlo Prime, Quantum yielded a lower background, but less signal for the GAPDH bands. This could be compensated for by using a longer exposure.
None
Rapid signal with lower background
Less signal intensity; cost ($245 for 100 ml)
Worth using with a good antibody and/or decent antigen amounts.
Product Name: WesternBright HRP SubstratesSupplier Page