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Antibodies That Work Again and Again and Again

Kameron Simpson1, Michael Browning2

  • 1PhosphoSolutions, Colorado Bioscience Park, 12635 E. Montview Blvd #213, Aurora, CO, 80045, USA.

Methods in Molecular Biology (Clifton, N.J.)
|February 11, 2017
PubMed

In the past few years significant concern has been raised about the quality and reproducibility of antibodies used in numerous scientific publications. In this chapter we discuss some of the biggest contributing factors to the "antibody problem" from both the commercial production side, as well as the end-users side. Specifically we argue that Western blot data should be used to provide a reliable initial indication of antibody quality, as well as a guide to distinguish between multiple offerings for antibodies to the same target. Secondly, we describe a set of best practices for antibody manufacturers to employ that will eliminate most of the variability in polyclonal antibodies. Taken together these proposals provide a way to significantly improve both the quality and the reproducibility of commercial antibodies.

Keywords:
Antibody specificityAntibody validationAntibody variabilityCell lysateLot-to-lot variationsPhospho-specific antibodiesRecombinant proteinsWestern blot

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