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. 2014 Aug 11;289(39):27019–27033. doi: 10.1074/jbc.M114.593780

FIGURE 9.

FIGURE 9.

Analysis of TRPM1 in mouse retina. A, retina lysates (∼50 μg) from wild-type (WT) or Trpm1−/− (KO) mice were subjected to SDS-PAGE and Western blotting for TRPM1 with mAbs 274G7 and 545H5, or actin antibody, followed by anti-mouse HRP secondary antibody, secondary antibody only, or biotinylated 545H5 (biot-545) followed by streptavidin-HRP. Endogenous antibody heavy chain dimers (Ab HC) are detected by the secondary antibody. B, detergent screen. WT retina lysate was incubated with the indicated detergent for 90 min at 4 °C followed by centrifugation at 100,000 × g for 1 h. Pellet (P) and supernatant (S) fractions were analyzed by SDS-PAGE and Western blotting with 274G7. C, retina lysates from WT and KO mice were solubilized with fos-choline-14 (Fos-14) and analyzed by BN-PAGE and Western blot with biotinylated 545H5 (left) or SDS-PAGE and Western blot with 545H5 or actin antibody (right). Fos-12, fos-choline-12; DM, decyl β-maltoside; DDM, dodecyl β-maltoside; OG, octyl β-glucoside.