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.
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 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 “Antibodies and westerns blots, the case of the missing
protein." This is better known as what happens when you use a poor lysis buffer.
