I want to select an antibody (calpain-1 antibody) for western blotting. While selecting an antibody I have some questions in my mind which need more clarification to proceed further. These are the questions and I seek the help of someone.
1) Which mab do I have to select (either polyclonal or monoclonal)?
2) Is there any effect of epitope on the overall outcome?
3) Which species do I have to select (for example if i want to check expression of calpain- 1 from rat tissue) and which mab do I have to select (mouse or rabbit mab)?
4) From where do I have to purchase the antibody (Santa cruz/ CST/ Abcam)?
Thanks in advance.
Hi, Lakshmi,
monoclonal antidy may have a nigh specficity but may have lower signal. If both is work well, it is better to use monoclaonal.
Of course, epitope is important, but epitope is much more important for immunolocalization,
For the company, it is up to you. All of them produce high quality antobody.
Good luck!
I would find a published antibody in a similar situation i.e western blotting rat tissue. There are pros and cons with both mono and polyclonal, but is unknown which is best until you try. It's impossible to predict. That's why I always try to find someone publishing it first. At the very least make sure the supplier has a photo of the western blot on the website. Be sure to check their western blot is detecting endogenous protein since a lot of the time they will only show detection in cells highly over-expressing the protein after transient transfection.
Hi Lakshmi,
Often the tech support of the suppliers can help you retrieve information about the performance of a certain product such as scientific references, in-house or customer data. There are lots of antibody suppliers, there may be more than the three you have mentioned that have the antibody. Several suppliers have reviews (Novus Biologicals, Abcam etc) where users can upload their own data. There are also testing programmes such as the Innovator award at Novus and the Abtrial at abcam to mention a few where you can get 100% reimbursement of the cost of the antibody if you are testing it in a new application so that you don't have to waste money. Good luck!
You can also do a search using the antibodyepedia.com to compare antibodies from different suppliers. The advantage of this site is that they will tell you the applications for which the antibodies were tested.
However, after you choose the best options, I would do as others already mentioned: search for publications, contact the tech support, etc.
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