Fig. 2.
Assessment of isolation efficiency and specificity of immunoaffinity purification. (a) A GFP-tagged bait protein was isolated (see Subheading 3.3.2) and eluted with the alternate detergent-based procedure. Ten percent of the following fractions are analyzed to assess tine efficiency of isolation of a GFP-tagged bait protein: FT flow-through; PE primary eluate; SE secondary eluate. The majority of bait protein was found in the primary eluate. (b) Immunoaffinity purification of endogenous Shp2 protein from 100. stage 40 Xenopus tropicals whole embryos. As a control for the specificity of the Shp2 antibody, a second immunopurification was performed in parallel using an anti-V5 antibody.