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. Author manuscript; available in PMC: 2008 Oct 8.
Published in final edited form as: Biochemistry. 2006 Jan 24;45(3):976–986. doi: 10.1021/bi051978h

Figure 5. Reversed-phase HPLC purification of [3H]Azicholesterol labeled fragments from tryptic digests of αV8-10, βV8-12, γV8-14, and δV8-11.

Figure 5

For αV8-10 (A), βV8-12 (B), γV8-14 (C), and δV8-11 (D) isolated from nAChRs labeled with 1.25 μM [3H]Azicholesterol in the absence of agonist, tryptic digests were fractionated by reversed-phase HPLC on a Brownlee Aquapore C4 column (100 × 2.1 mm) as described in the Experimental Procedures. The elution of peptides was monitored by absorbance at 210 nm (solid line) and elution of 3H by scintillation counting of aliquots (25 μL) of each 500 μL fraction (●). The point of elution of the M4 peptide based on previous work (and results from Fig. 6) is indicated with an arrow. Based upon recovery of radioactivity, >90% of the material was recovered from the HPLC column. HPLC fractions 38-41 (A) or 37-40 (B-D) were pooled for sequence analyses (Figure 6).