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The image is totally clean so could I double check? I blot the same membrane for anti-p21 and I obtained a band to the correct size
have you used 400 cells/ well or 50- 90ug protein/ lane? The image that I have attached to the e-mail represent different µg of whole lysate for lane. In another gel I tried to load 400.000 cell/line as protocol that I used in my lab.
- Did you observe same clean membrane after trying 3 times? yes
- Is the secondary antibody fine? Have you tried "loading control" with the same lysates? the secondary antibody is correct and I load the same lysate on another gels and I blotted the membrane with other antibodies.
- Is it true no protease inhibitors were added? Could you try after adding a PI cocktail? the Laemmli buffer does nor require the addition of protease inhibitors.
I would like to test another batch of this antibody and if I have the same results I buy another antibody.
Thank you for your attention on this matter.
Asked on Oct 17 2012