Green and red IgG

Antibodies are extremely valuable research tools, however producing an antibody that works can be very challenging. Both bad antibodies and bad antibody technique can lead to frustration for researchers.

Contributed by Mike Browning

Production of good antibodies is time consuming and difficult, and not all antibodies will work in all assays. 

Synapsin IHC

Antibodies that work are extremely valuable research tools. However, the search for such antibodies is made quite difficult by two different problems. The first problem, “the good antibody problem” is that production of a good antibody is a difficult, time consuming process; and many researchers and antibody companies do not possess either the patience or the skill necessary to make good antibodies. The second problem, “the bad technique problem” is that a good antibody may not “work” in all of the various antibody-based assays. This may occur because the antibody cannot detect its target in a particular type of assay (e.g. the antibody target may become denatured in formalin fixation, thus formalin fixation is a bad technique for this antibody).  Alternatively, incorrect technique (AKA bad technique) in an assay may interfere with the antibody binding to its target.

Antibody performance can drastically affect research outcomes.

I have been working with antibodies for quite some time and I have had many opportunities to observe antibodies that do and do not work. For my students who used antibodies that worked, the outcome was often the development of important new insight into a specific protein’s role in normal or disease function. In contrast, months of frustration and false leads were usually all that resulted when my students used antibodies that did not work. In this blog I will attempt to discuss how to address both the good antibody problem and the bad technique problem so that more antibodies will work. 

I hope this blog will bring together many scientists who work with antibodies and provide a forum for support and discussion. 

I am hoping that this will become a collaborative project and I invite anyone who cares about antibodies to contribute to the discussion by offering tips, advice and/or topics for discussion. I look forward to talking with you on this blog in the coming months.My next topic will be on a bad technique problem and is titled “1% SDS is the Lysis buffer of Choice for Western Blots, or The Case of the Missing Protein.” This is better known as what happens when you use a poor lysis buffer.

Read next:

Antibodies that DO NOT Work in Western Blot

All Purified Antibodies are NOT Created Equal

Antibody Variability

Antibody Dilution Recommendations - What you need to know!

Further Reading:

SDS-PAGE Demystified – The Science Behind All Those Bubbles!

Antibodies in Western and IHC

Antibody Variability

Are Monoclonal Antibodies Really More Specific?

1% SDS is the Lysis Buffer of Choice for Most Western Blots

Our full blog

More educational resources


Antibodies in western and ihcPhosphosolutions