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. 2003 Mar 19;100(7):3995–4000. doi: 10.1073/pnas.0230488100

Figure 1.

Figure 1

Characterization of anti-PI4KIIα Abs. (A) HEK293 cells were transiently transfected with myc-tagged PI4KIIα and analyzed by Western blotting. Both anti-myc and anti-PI4KIIα Abs recognized a major band at ≈55 kDa and a minor band at ≈66 kDa in transfected cells. (B) Triton X-100 extracts of transfected and mock-transfected cells were immunoprecipitated (IP) with either anti-myc or anti-PI4KIIα rabbit polyclonal Abs. Immunoprecipitates were then subjected to Western blotting (WB) with either mouse monoclonal anti-myc Abs or biotin-labeled anti-PI4KIIα Abs followed by streptavidin. (C) A Triton X-100 extract of rat brain was immunoprecipitated with anti-PI4KIIα or anti-PIPKIγ Abs. The immunoprecipitates (IP) were subjected to a PI kinase assay, and the products were analyzed by TLC and autoradiography. (D) Anti-PI4KIIα immunoprecipitates generated from rat brain Triton X-100 extract were analyzed by the PI kinase assay in the presence of the indicated compounds. (E) TLC analysis of reaction products generated by anti-PI4KIIα immunoprecipitates from Triton X-100 brain extracts in the presence of several synthetic PI substrates. X denotes an unidentified spot.

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