I have continually had issues with bad looking western blot images, specifically black dots and smears on the nitrocellulose membrane. Does anyone know what could be the cause of this? I have read that this could be due to the blocking buffer - I am using a 1:1 ratio of TBS (with 0.1% Tween) and Intercept Blocking Buffer. If this is a blocking buffer issue, are there better blocking buffer recipes I should use?
Thanks in advance!
Please use fresh blocking buffer, increase blocking time. Use tweezer for handling membrane.
Hi Alex! I do the blocking for 1 h at RT in 5% skim milk in TBS (without Tween-20). Regarding the smear, if you work with homogenates I suggest you to do a short sample sonication prior to loading, in order to avoid the smearing.
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