Advansta
AdvanBlock-Chemi blocking solution
R-03726-E10
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I have used AdvanBlock-Chemi blocking solution for blocking both western blots and immunofluorescence staining. For westerns, one can go from blocking to detection in about 3.5 hours. I have better results and less background than with classical drymilk blocking solutions. Because the solution is also an antibody binding enhancer, one can use less primary and/or secondary antibodies.
Western blotting
Standard western blot with whole cell lysates on PVDF membrane
Reagent protocol calls for three 1 hr incubation steps: blocking; primary and then secondary antibodies. Washes are two rinses with TBS-T, followed by three 5 min washes. If time is an issue, one can either block or incubate with primary antibodies overnight in the cold. I use 3 ml of reagent in a rotating 50 ml tube.
One does not have to change the reagent after blocking step. Just add primaries. The reagent also works for IFF, but an overnight incubation with primary antibodies is needed.
As can be seen in the figure, the blot probed using Advanblock had much higher sensitivity and signal intensity with two different antibodies.
N/A
Advansta has 30 ml of the reagent available for free to test. That will allow for some 2-5 blots, depending upon your system.
Rapid results; one liter of reagent will last a long time!
None so far!
Outstanding reagent