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. 2003 Jun 18;551(Pt 3):751–763. doi: 10.1113/jphysiol.2003.040014

Figure 7. Co-immunoprecipitation of KV1.2 and 1.5 from rat cerebral arteries.

Figure 7

A, stably expressed KV1.5 in HEK cell membranes (1 μg) detected by anti-KV1.5 blot as a positive control. B, anti-KV1.5 immunoprecipitate from cerebral arterial membranes (200 μg) blotted with anti-KV1.5 revealed a 75 kDa band, indicating successful immunoprecipitation of KV1.5. C, stably expressed KV1.2 in HEK cell membranes (1 μg) detected by anti-KV1.2 blot as a positive control. D, native KV1.2 in cerebral arterial membranes (8 μg) detected by anti-KV1.2 blot as an ˜80 kDa band. E and F, anti-KV1.5 immunoprecipitate from 50 μg and 30 μg of two different cerebral arterial membrane preparations. An anti-KV1.2 blot revealed the same ˜80 kDa band in both arterial preparations, indicating successful co-immunoprecipitation of KV1.2 and 1.5. G, preadsorption of anti-KV1.2 with 1 μM of its antigenic competing peptide (+CP) abolished the 80 kDa-immunoreactive band detected in anti-KV1.5 immunoprecipitate. Anti-KV1.5 immunoprecipitate was from 30 μg of the same pool of arterial proteins as used in F. Results using cerebral arteries were verified in a minimum of four different protein preparations isolated from ˜60 rats.