Rosano C L, Braun C B, Hurwitz C
Clin Chem. 1987 Mar;33(3):398-400.
The radial immunodiffusion assay overestimates the C1q in serum. Here we describe a convenient, accurate procedure for measuring C1q in 250 microL of dialyzed serum. This method is based on our previous findings that all C1q in serum precipitates with the euglobulin fraction and that all other serum proteins containing hydroxyproline are excluded from this fraction. Because C1q is 4.3% hydroxyproline, the concentration of C1q in serum can therefore be calculated from the hydroxyproline content of the euglobulin fraction. The procedure, all done in the same tube, consists of precipitating the euglobulin fraction, digesting it with HClO4, and converting hydroxyproline to the corresponding pyrrole, which is extracted with toluene and measured by absorbance at 560 nm.
单向免疫扩散试验会高估血清中的C1q含量。在此,我们描述一种简便、准确的方法,用于测定250微升透析血清中的C1q。该方法基于我们之前的研究发现,即血清中的所有C1q都与优球蛋白组分一起沉淀,而所有其他含羟脯氨酸的血清蛋白都被排除在该组分之外。由于C1q含4.3%的羟脯氨酸,因此血清中C1q的浓度可根据优球蛋白组分中的羟脯氨酸含量来计算。该操作均在同一试管中完成,包括沉淀优球蛋白组分、用高氯酸消化、将羟脯氨酸转化为相应的吡咯,再用甲苯萃取并在560nm处通过吸光度进行测定。