Response: ‘Antibody crossreactivity between the tumour suppressor PHLPP1 and the proto‐oncogene β‐catenin’

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Lobert and co‐workers [1] reported a consequential flaw in the immunoreactivity of the antibodies A300‐660A and IHC‐00382 to PH domain leucine‐rich repeat protein phosphatase 1 (PHLPP1) manufactured by Bethyl Laboratories, Inc. (Montgomery, Texas, USA). The flaw is the crossreactivity of these preparations with β‐catenin. This response is to acknowledge the validity of the report of Lobert and co‐workers, to report that testing at Bethyl Laboratories has confirmed their findings and to report production of antibodies to the carboxyl‐terminus of PHLPP1 with less than 3% residual crossreactivity with β‐catenin when compared with crossreactivity of A300‐660A to β‐catenin. The antibodies A300‐660A and IHC‐00382 had a common means of production in which a peptide (LPDYYDTPL) representing the C‐terminus of PHLPP1 was conjugated to a carrier protein and used as an antigen for the production of hyperimmune sera in rabbits. Subsequently, antibodies to the peptide were purified by using the peptide conjugated to beads of agarose. The affinity‐purified antibodies were tested in immunoblotting and immunoprecipitation. Having found that the preparation of antibodies was useful for immunoblotting and immunoprecipitation, the preparation was offered as part of Bethyl's line of products as product number A300‐660A. Subsequent to testing for usefulness in immunohistochemical staining, a portion of A300‐660A was reformulated to produce product number …