Zingde S M, Shirsat N V, Gothoskar B P
Anal Biochem. 1986 May 15;155(1):10-3. doi: 10.1016/0003-2697(86)90216-2.
A procedure is described for obtaining peptide maps from microgram quantities of protein in gel bands, after cleavage at the methionyl peptide bonds with vapors of acidic cyanogen bromide (CNBr). Absence of direct contact of the gel pieces with CNBr eliminates the need for extensive equilibration of the gel piece to remove CNBr prior to electrophoresis. The milder conditions lead to partial cleavage of the proteins, yielding larger peptides and thereby reducing the risk of peptide loss during the postelectrophoresis procedures. The "fingerprints" obtained are reproducible and independent of an eightfold change in CNBr concentration.
本文描述了一种方法，用于在酸性溴化氰（CNBr）蒸汽作用下，在甲硫氨酰肽键处裂解后，从凝胶条带中的微克量蛋白质获得肽图。凝胶片段不直接与CNBr接触，无需在电泳前对凝胶片段进行广泛平衡以去除CNBr。较温和的条件导致蛋白质部分裂解，产生较大的肽，从而降低了电泳后程序中肽损失的风险。获得的“指纹图谱”具有可重复性，且与CNBr浓度八倍的变化无关。
