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. 1998 Jan 15;18(2):610–624. doi: 10.1523/JNEUROSCI.18-02-00610.1998

Fig. 2.

Fig. 2.

Detection of hair bundle PMCA with the 5F10 antibody. A, Immunoblot detection of proteins of 140 and 170 kDa in isolated hair bundles and residual macula, the sensory epithelium remaining after hair bundle isolation, with anti-PMCA monoclonal antibody (5F10). Hair bundles, From 21 frog sacculi (∼42,000 bundles in agarose gel); Agarose, equivalent amount of agarose gel; Residual macula, 0.5 saccular equivalents; RBC PMCA, purified human erythrocyte PMCA (5000 pg). B, Immunoprecipitation of PMCA with 5F10 from a Triton X-100 extract of purified, biotinylated hair bundles. Equal fractions of the Triton X-100 bundle extract, flow-through solution from the 5F10 anti-PMCA antibody precipitation, and 5F10 SDS eluate were loaded. C, Immunoprecipitation of 140 and 170 kDa calmodulin binding proteins with 5F10 from purified, biotinylated hair bundles. Calmodulin binding proteins were isolated first from a Triton X-100 extract of purified, biotinylated hair bundles, using calmodulin-agarose; then the EGTA eluate from the calmodulin-agarose was used for precipitation of PMCA with 5F10. Equal fractions of each sample were loaded.