The Advansta team hopes that the holiday season gave you all an opportunity to relax and recharge. A new year brings with it opportunities for new beginnings. Now that everyone is back to work we hope that there are some exciting new experiments on the horizon. If you are like us, every great experiment starts with selecting the right antibody. So much of our success depends on the performance of this one critical reagent, so it is well worth the time and effort to select the right one for the job. Here are some questions that we ask ourselves before we buy our antibodies.
1. What techniques will I need to perform with this antibody? This is an important question to start with because primary antibodies do not recognize native and denatured proteins the same. If you need to run an ELISA and a Western with the same antibody, make sure to choose one that has been validated for both applications. In addition to the convenience of only having to buy one antibody, your data will be more consistent and easier to interpret if you use the same antibody across all experiments. For help making your selection, check out the product information section on the manufacturer’s website. This section typically lists the compatible applications.
2. Has the antibody been validated? Many suppliers have validation data for each antibody posted on their website. Although this is very helpful, we suggest taking it a step further and confirming that data independently. Luckily, there are databases available such as CiteAb, which ranks antibodies by citations to help you select antibodies that have been successful in past research papers. When it comes to Western blotting, my personal lucky number is 15. Those antibodies with at least 15 references tend to require very little optimization, making blotting a breeze.
3. Is the supplier reliable? Unfortunately, being charged $400 for an antibody doesn’t mean that you are getting a quality product. In fact, many suppliers do not even produce all of their own antibodies. They fill their catalog with antibodies purchased from other suppliers since it is often too challenging to produce all the required antibodies internally. The biggest risk of purchasing from an unreliable supplier crops up when it comes to long term projects. If you happen to pick a poor selling antibody, there is no guarantee that you will have a consistent supply and it is not uncommon for a company to ditch an antibody that isn’t selling well and replace it with another. Additionally, companies that are not reliable may have lower quality control standards, resulting in high lot-to-lot variability which will wreak havoc on your experiments.
Screening primary antibodies can be extremely expensive, so don’t be afraid to look for a good deal. While some companies may provide you with a small sample free of charge, others are now selling smaller aliquots for evaluation. If you are short on funds, you can also try to barter validation data in exchange for a trial sample. Another tip is to keep an eye out for companies that guarantee the performance of their antibodies in validated applications. That way, if the antibody fails to perform you can work with them to resolve the issue or find a replacement that better suits your needs.
We know that choosing a primary antibody can be an onerous task, but with some careful research and a little bit of leg work you can increase your odds of finding one that’s right for you!