How to get low molecular weight bands (i.e. TNF-alpha, TGF-beta, IL-1beta) by western blot
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
Also standardize your transfer time and current conditions. Preferably use 0.22 micron pore size membranes.
mainly transferis very important ,
Transfer should be done for longer time