Dear Colleagues
I have a colleague who is not expecting an antibody in her lysate to give expression of a protein at about 23kD (owing to gene methylation)
However, the antibody does give bands of a much higher Mr. What is more, there is no positive control on this blot which (would validate the antibody)
Am I right in thinking that unless you have a positive control on the blot which validates the antibody i.e. produces a band of 23KD then you cannot infer that other lysates do not express (by absence of that band). By extension, there is little point in stripping this blot and re probing with actin (?)
Rather a sensible course of action would be to repeat the blot and include a positive control; and then strip and re probe with actin (as a loading control)