I am looking for the best Western Blot protocol in www.abcam.com but I know working with monoclonal is harder than polycolonal.
Could you please tell me your experience?
There is not specific protocol for western blotting for N-cadherin only you can use same protocol as you used for any other, it doest matter either you use monoclonal or polyclonal antibody, what is necessary is you should know about the molecular weight for that specific protein, beside that you need to use secondary antibody produced in same species i.e. if you are using the primary antibody produced in rabbit then you should use secondary antibody of rabbit as well. Recentally I used the N-cadherin abtibody of abcam in the concentration of 1:1000... the gel used for western blotting was 8% gel and molecular weight for N-cadherin was 135 Kd..
I agree there is definitely no diff either you use a mono or polyclonal ab, except that most likely the mono is more pure ( prot G or A purified in general) so it generally gives you a cleaner blot, less risk of unspecific band since it recognizes only one epitope.
dont waste your time looking for best protocol as there is none and yep no such thing as a clear fast rule of monoclonal antibodies being harder to work with than polyclonal. Antibodies can either work great or be pure rubbish and that is the same for both monoclonal and polyclonal.
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