If you’re working in the life sciences, chances are you’re going to be using antibodies in your research. You’ll probably be asked by your supervisor to find suitable antibodies, so I, Dr Andrew Chalmers, founder of CiteAb, have set out some top tips to get you started in making a good antibody buying decision.
When you are choosing your antibody, you need to take your time.
Not all antibodies will be suitable for your experiments so taking a bit of time choosing an antibody can stop you wasting many days or even weeks at the bench. This is especially true with new researchers – you don’t want to end up blaming yourself for a failed experiment, when in reality it is the antibodies fault!
Use an antibody search engine.
The reason we set up CiteAb was because as a researcher I got frustrated with the time and money wasted by my lab when we were trying to find good antibodies. The right antibody might not come from a well known supplier so its worth using an antibody search engine to aid your search. I would obviously love it if you used CiteAb 🙂 We list over 2 million antibodies and rank by citations, which means companies can’t pay to rank higher, so we have to be a good place to start!
Independent validation is crucial.
We believe the best way to find an antibody that works is to see that it has already been used by an independent lab and the results published. We can help you find these citations and you can then check to see if the data looks good. Antibody reviews and independent validation by other companies can also help give you confidence, but just like citations you should look at the detail and quality of the data provided before making any decisions. It is also worth remembering that you will still need to validate the antibody in your lab.
Is the antibody suitable for your application?
Just because an antibody has been successfully used in one application does not mean it will work for all applications.
Does the antibody work with the species you are using?
The majority of antibodies are raised against human proteins so if you are not working with human samples be careful to check the antibody will cross react with the species you are using.
Monoclonal vs polyclonal.
My advice is you should go with the most validated antibody, but if you are lucky enough to have two well validated antibodies to chose between, then monoclonal antibodies may suffer from less batch to batch variability than polyclonals (although you can’t assume no variability). Polyclonals from large animals like goats may also have less batch to batch variability.
Check your host species.
Its very easy to use the wrong secondary antibody and a blank western blot is so painful, so make sure you check which species your primary antibody is raised in. This is especially important if you are doing double staining and need two different species of secondaries. If using goat antibodies it is also worth checking if your secondary antibodies are raised in goat and may cross react.
Several companies may sell exactly the same antibody.
A manufacturer may license their antibody to many companies, so do not buy five antibodies to test without making sure they are actually different. You can do this by asking where the antigen is found within the protein or checking they are raised in different species. With common monoclonals that are supplied by many companies you might also consider shopping around for the best supplier of a clone.
If the antibody does not work ask an expert.
Do the obvious checks to make sure you did not forget a key reagent and then ask for advice – don’t just keep doing the experiment again. Find somebody in your department or email an author who has used the antibody. Also contact the company for help, see below.
If the antibody does not work contact the company.
You should feel free to contact the company for advice and ultimately a refund. The suppliers really should be helpful, many pride themselves on their customer service, and if they are not you can remember that for next time you buy an antibody!
And of course once you get your great data and write it up, keep our development manager Matt happy and cite the antibodies properly – we have a guide here to help you do that!
Good luck for these first few weeks of your PhD from the whole team here at CiteAb. If you have any questions about finding antibodies to use in your work do comment below and we’ll do our very best to help you out.
– from Andrew the CiteAb team