title: Antibodies That Work Again and Again and Again.
abstract: 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.
authors.affiliation: PhosphoSolutions, Colorado Bioscience Park, 12635 E. Montview Blvd #213, Aurora, CO, 80045, USA.
authors.forename: Kameron
authors.initials: K
authors.lastname: Simpson
authors.affiliation: PhosphoSolutions, Colorado Bioscience Park, 12635 E. Montview Blvd #213, Aurora, CO, 80045, USA. MDB@phosphosolutions.com.
authors.forename: Michael
authors.initials: M
authors.lastname: Browning
mesh_terms: D000818:Animals; D000906:Antibodies / Q000276:immunology*; D000911:Antibodies, Monoclonal / Q000276:immunology; D000918:Antibody Specificity / Q000276:immunology*; D015153:Blotting, Western; D005455:Fluorescent Antibody Technique; D006801:Humans; D010750:Phosphoproteins / Q000276:immunology; D011994:Recombinant Proteins / Q000276:immunology; D015203:Reproducibility of Results
chemical_list: D000906:Antibodies; D000911:Antibodies, Monoclonal; D010750:Phosphoproteins; D011994:Recombinant Proteins
keywords: Antibody specificity; Antibody validation; Antibody variability; Cell lysate; Lot-to-lot variations; Phospho-specific antibodies; Recombinant proteins; Western blot