Multiple bands on a Western blot and solutions for western blot background
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Multiple bands on a Western blot and solutions for western blot background (archive)
A FAQ from Synaptic System on problem of many bands on a western blot.
Beside solutions provided above, some other considerations may be helpful for eliminate multiple bands in western blot.
Some solutions for high background of western blot (multiple bands):
1. Increase the washing stringency (longer wash, faster shaking).
2. Increase the blocking reagent concentration in both primary and secondary antibody inclubation steps.
3. Load less proteins per lane.
4. Decrease concentrations of primary and secondary antibodies.
5. Try a less sensitive detection method.
Last update 16-Mar-2005, Rating Good of 2 votes.
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I had unspecific bands on my blot which made it difficult for me to dentify my protein. all bands were very faint, I am afraid if i increase the dilution of my antibody that I will not be able to visualize my protein
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try a different larger dilution of antibody in the incubation buffer
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