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I would like to test the Anti-HPRT Antibody[EPR5298] ab133242. I want to use this rabbit monoclonal antibody in western blot. I am using mouse brain tissues.
The main reason I want to join the Abtrial is I will use another rabbit polyclonal to 5HTR2A antibody (ab16028) in the same blot membrane. I don't know if my experiment will be messed up since I use two primary antibodies both from rabbit in one membrane.
Please give me some suggestions base on my experimental situation!
Asked on Mar 26 2014
The AbTrial is not applicable to tests of ab133242 for staining western blots of mouse samples, as this combination (western blotting and mouse) has already been tested. Our Abpromise guarantee will apply, though, as it covers western blotting of mouse samples with ab133242. For details of our guarantee, please see the page at www.abcam.com/abpromise .
Your plan to stain the same blot with two different antibodies will not be difficult. Most people attempt this by cutting the membrane in half. The two proteins you are trying to detect are 25 kDa (HPRT) and 53-60 kDa (5HT2A receptor). So, if you cut the membrane at about 40 kDa, using pre-stained molecular weight markers to guide you, you can stain each half separately, one half for each antibody.
If you do not cut the membrane, and stain it with both antibodies, then you will not know which bands were produced by which antibody.
Tom RuyleAbcam Scientific Support
Answered on Mar 26 2014