western blot

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arun prasath
arun prasath's picture
western blot

Dear friends,

How to get low molecular weight bands (i.e. TNF-alpha, TGF-beta, IL-1beta) by western blot

protoldo's picture
Hola , Run a 15% PAGE

Hola , Run a 15% PAGE,Transfer it without stain to a nitrocellulose or PVDF membrane, Block with 5% powdered skimmed milk in PBS 0.1% tween20 incubate with the specific primary antibody in milk solution, (cutting the lanes of each different sample or a mix of the first ab. if there aren´t interferences, Wash with PBS tween and incubate with the secondary ab. coupled with the fluorofore if you want reveal by fluorescence or HRP if you want see the bands by chemiluminescence in autoradiography. Wash and develop. There are lots of similar method in the net. Good luck

sharmakaushikp's picture
Also standardize your

Also standardize your transfer time and current conditions. Preferably use 0.22 micron pore size membranes.

Rajeshwari patel
Rajeshwari patel's picture
mainly transferis very

mainly transferis very important ,

Transfer should be done for longer time