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A Guide: how to choose the right antibody.

Five tips for choosing the perfect primary antibody

Written/Edited by Dr. Ryan Robinson, PhD

Alright - you’ve got your funding, you've got your equipment, and now you have the perfect experiment planned to get that result you just KNOW is going to define your career, or at least keep your PI happy and off your back for a little while. Now all you need is the perfect antibody – so…. how do you find it?

At antibodies-online, we understand that finding the right primary antibody can be a daunting task. With a catalog that contains more than 1,400,000 primary antibodies from hundreds of worldwide suppliers chances are good that we can help you find the right reagents, and with the industry’s fastest and most innovative search we make it easy to locate them. Here are a few tips to help you minimize the amount of time you spend searching for the right antibody so that you can get your experiment up and running as quickly as possible:

1. Plan ahead

It’s important to remember the 5 Ps (Proper Planning Prevents Poor Performance) in any field, but perhaps nowhere do they have a larger impact than in science. The most important phase of any experiment is the planning phase.

Chances are good that you already have the basic details of your experiment worked out: What target you want to look for (antigen), what species you want to work with (reactivity), and what sort of experiment you want to perform (application). You may also have some of the more specific details planned, e.g., which epitope you'd like to target, whether you would rather use a monoclonal or polyclonal antibody, or if you'd prefer to use directly conjugated primary or employ a secondary antibody. You can start searching for compatible antibodies right away by using our search function and innovative smart-filters to help you rapidly pare down hundreds of thousands of options and find just the right product.

If you're having trouble finding the right reagents to help you complete the experiment in the way you planned, you may want to put some thought into a less-common consideration. Take a moment and ask yourself: "Is my method the best way to answer this question?" It's very easy to get tangled up in the familiar, and to limit yourself to the tools and techniques that you know and have used before. Remember that creativity, innovation, and forward thinking are all core pieces of the scientific process. There may be a better way to solve this problem, think outside the box and don’t become irrevocably committed to one approach or technique!

Researchers often become intently focused on specific methods and eschew alternate approaches in favor of tried-and-true techniques, but a non-conventional solution may offer a significant improvement over the traditional approach. For example, our GFP Catcher allows a researcher to quickly and cleanly purify GFP-tagged fusion proteins while avoiding many of the difficulties and frustrations associated with traditional IP techniques.

2. Know the details

Choosing the perfect antibody is a lot like performing any experimental technique - the devil is in the details. The more you know about the target that you are looking for, the sample you're looking at, and the nature of the experiment you have planned, the easier it will be to find the right reagents.

You might know that you want to stain a human sample, but do you know the source? You may need a different antibody for performing IHC on a than you would when performing IHC on a or performing immunostaining on .

WNT2 antibody  (AA 221-320) WNT2 antibody  (AA 221-320) WNT2 antibody (AA 221-320) (ABIN762896)
  • Paraffin Embedded Human Brain Section Wnt2 Staining
  • counterstained with hematoxylin (blue)
  • In stock, fast delivery
CD163 antibody CD163 antibody CD163 antibody (ABIN111878)
  • Frozen Human Placental Staining
  • Section CD163
  • In stock, fast delivery

Knowing the strengths and limitations of your primary antibody can help you get the best results from your experiment. Both of the antibodies above produce stunning IHC images, albeit on different tissues and via slightly different methods.

3. Choose the right controls

It is a universally accepted axiom that no experiment is a failure unless you learn absolutely nothing from it. When planning an experiment, having the right controls in place is often the defining factor that distinguishes success from failure. In addition to your own context dependent controls, most common experimental techniques require specific controls to help distinguish positive and negative results from background or user error.

For example, Western blots should always include a loading control (usually some form of broadly expressed housekeeping protein) and IHC/IF experiments should include one or more negative control samples to help distinguish signal from background noise. If you need help establishing the right control for your experiment ask one of our technical support scientists.

Custom Antibody Services

Custom Antibody Services

Benefit from combined expertise of antibodies-online and our Partner Rockland Immunochemicals, Inc. Collaborative, experienced, and conducted completely in our own facilities, our custom antibody production services can drive your project from the design and synthesis of your antigen through to production and characterization of your antibody.

Benefit from combined expertise of antibodies-online and our Partner Rockland Immunochemicals, Inc. Collaborative, experienced, and conducted completely in our own facilities, our custom antibody production services can drive your project from the design and synthesis of your antigen through to production and characterization of your antibody.

Learn more

4. Learn the protocol, and set a standard for reproducibility

If proper controls are the most important distinguishing factor between a successful experiment and a failed one, it is only because reproducibility is a close second. Sadly, reproducibility and scientific rigor have fallen by the wayside in favor of rapid progress. Recent evidence suggests that, in some fields, fewer than 30% of published studies can be replicated by an independent laboratory.

Independent Validation Initiative

antibodies-online is doing its part to help reverse this trend. We started the Independent Validation Initiative(IVI) to actively approach the problem. The IVI was launched in July 2013 to independently validate commercial antibodies and ELISA kits. Our aim is to increase the transparency of product quality in the antibodies and ELISA kits market by providing exhaustive background information on how the product was applied in the experiment, while also allowing our customers to review results.

You can help too! We encourage qualified scientist to become part of the IVI by proposing a validation experiment for a recommended application. Become a Validator

Carefully reviewing the manufacturer's recommended experimental protocol when choosing a primary antibody can help ensure that the product you are choosing is compatible with the experiment you intend to perform. Deviations from a standard manufacturer approved protocol are occasionally permissible, but should only be applied when absolutely necessary and should always be carefully documented. Closely following a well-documented protocol can also help immensely when attempting to troubleshoot any unforeseen difficulties that may occur during the course of your experiment.

If you need help finding a specific protocol, our scientific support staff can provide you with a manufacturer approved protocol for any product in our catalog - just ask!

5. Don’t be afraid to ask for help

Science is all about asking questions, and antibodies-online is always happy to help you find the answer. Whether you are just starting to plan your project, or you already know exactly what you are looking for, the antibodies-online scientific support staff can help you find just what you need to complete your experiment. Our service doesn’t stop once you’ve found the right product either. Our experienced team of support scientists will help you apply your product and troubleshoot any issues you encounter along the way. You can contact a support scientist by emailing us directly:

Ryan Robinson
Dr. Ryan Robinson, PhD
Scientific Content & Communications Manager, antibodies-online alumn

Offered pre- and post-sales consultation and support to antibodies-online customers. Provided troubleshooting and technical assistance with proteomics and genomics experiments. Composed and distributed cross-channel digital marketing campaigns.

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