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. 2004 Feb 4;24(5):1236–1244. doi: 10.1523/JNEUROSCI.4512-03.2004

Figure 6.

Figure 6.

Immunoblots and immunoprecipitations. A, B, Immunoblot analysis. Membrane proteins (100 μg) from rat muscle, sciatic nerve, spinal cord, and brain, and HeLa celllysates (10 μg) were separated by electrophoresis and immunoblotted for KCNQ2 (A) or KCNQ3 (B). Bands corresponding to the molecular mass of KCNQ2 and KCNQ3 expressed in HeLa cells (∼97 kDa) were detected in both spinal cord and brain. KCNQ3, but not KCNQ2, was detected in sciatic nerve membrane. C, Immunoprecipitations of KCNQ2 and KCNQ3. Rat optic nerve and hippocampal membranes (200 μg) were immunoprecipitated for KCNQ2 and KCNQ3 and then immunoblotted with KCNQ2 or KCNQ3 antisera. KCNQ2 and KCNQ3 were detected in both samples. MW markers are shown on the left (in kilodaltons). D, E, Coimmunoprecipitations of KCNQ2 and ankyrin-G. Rat spinal cord membranes (200 μg) were immunoprecipitated for KCNQ2 or ankyrin-G and then immunoblotted for ankyrin-G (D) and KCNQ2 (E). A ∼97 kDa isoform of ankyrin-G was pulled down by the KCNQ2. The ankyrin-G antiserum pulled down multiple ankyrin-G isoforms, including the ∼97 kDa isoform. KCNQ2 (asterisk) was immunoprecipitated by both the ankyrin-G and the KCNQ2 antisera. MW markers are shown on the left (in kilodaltons). In E, the immunoblot for KCNQ2 is shown for two film exposure times: 3 min (3′) and 10 min (10′).