I need to do immunoblot to check protein expression in and I hpoe to use mouse anti-myc antibody. I found two types of antibodies, myc monoclonal antibody and myc polyclonal antibody. Which type will be the suitable antibody for my work? How could I decide it?
Polyclonal antibodies are more robust and gives higher detection as they bind to multiple epitopes of the same antigen, so they detects even lower amount of antigen. They are also more stable as compared to monoclonal antibodies. However, The Monoclonal antibodies have quite high specificity for a particular epitope, therefore they are more advantageous in quantification experiments. Also chances of cross-reactivity goes down when we use monoclonal antibodies. Now, you can decide upon the requirements of your experiments, which would be suitable.
Polyclonal antibodies are more robust and gives higher detection as they bind to multiple epitopes of the same antigen, so they detects even lower amount of antigen. They are also more stable as compared to monoclonal antibodies. However, The Monoclonal antibodies have quite high specificity for a particular epitope, therefore they are more advantageous in quantification experiments. Also chances of cross-reactivity goes down when we use monoclonal antibodies. Now, you can decide upon the requirements of your experiments, which would be suitable.
I agree with Arun's answer. However, I suggest to find suitable references which mention these antibodies and decide on the same. And also, the manufacturers or sellers generally mention the application compatibility of these antibodies. If I am blind and can not decide, I go for polyclonal antibodies (look the published data if any for single band without any nonspecific binding on the manufacturer's website). Good luck
I also agree with the previous answers.
Both types of antibodies have their pros and cons. It depends on your needs: if you want to quantitate your signal later on and want to exclude any possible cross-reactivity, go for the monocolonal one; if you expect only low expression in your samples, go for the polyclonal one.
Some vendors also provide free or low-cost samples for testing. This might be an option if you feel unsure.
Yes, I agree with the above answers. Additionally, it depends on the starting material used as well. If you are using cell extracts or very low protein amount, polyclonal antibody would be a better bet due to the afore-mentioned reasons. Also, while choosing a western blotting antibody, also check which fragment(s), your antibody recognizes and choose wisely the one closest to your interest. I recommend Abcam, but be careful about the vial size (it normally is pretty less, but works at really high dilution as well).
Labome has reviewed over 800 articles, and compiled a list of antibodies used in those articles. You are welcome to check the results here https://www.labome.com/review/gene/human/c-Myc-antibody.html . You can follow the link to see the actual articles.
I agree with the points mentioned by others. In our lab, the anti-Myc antibody (9E10, mouse monoclonal IgG1) from Santa Cruz was used routinely. It detected Myc-epitope tagged recombinant proteins that were produced using the baculovirus system, and worked very well in Western blotting.
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