Mitchell J P, Van der Ploeg M, van Duijn P
Histochemistry. 1981;73(2):211-23. doi: 10.1007/BF00493021.
A comparison has been made between dinitrofluorobenzene (DNFB) and Naphthol Yellow S (NYS) as protein stains in combination with the pararosaniline-SO2 Feulgen procedure. Chicken erythrocytes were used as test cells. Cytophotometric measurements were made using a Zeiss scanning stage cytophotometer coupled to a PDP 11/10 minicomputer using the BICOSCAN program to obtain values for protein per cell, protein per "nuclear area' and DNA per nucleus. With 5N HCl as the Feulgen hydrolysis agent, DNFB staining, applied before the Feulgen procedure, was found to be unaffected by hydrolysis conditions required to give optimum Feulgen staining and showed only small losses after longer hydrolysis times. On the other hand measurements of NYS staining, of necessity applied after the Feulgen procedure, seem to be susceptible to the duration of Feulgen hydrolysis. This susceptibility is probably due to the interaction of the DNA phosphates with the basic amino acid residues, potential binding sites for NYS. Since the degree of this interaction may be variable, it is argued that NYS binding will measure the available basicity of proteins at the time of staining but no specific protein fraction. DNFB binding is unaffected by DNA-protein interactions and therefore can give a more reliable measure of "nuclear' protein, particularly in conjunction with Feulgen-DNA measurements.
已对二硝基氟苯（DNFB）和萘酚黄S（NYS）作为蛋白质染色剂与对品红 - SO₂ 福尔根程序联合使用的情况进行了比较。以鸡红细胞作为测试细胞。使用与PDP 11/10小型计算机相连的蔡司扫描台细胞光度计，通过BICOSCAN程序进行细胞光度测量，以获得每个细胞的蛋白质值、每个“核面积”的蛋白质值和每个细胞核的DNA值。以5N盐酸作为福尔根水解剂时，发现在福尔根程序之前应用的DNFB染色不受产生最佳福尔根染色所需水解条件的影响，并且在较长水解时间后仅显示出少量损失。另一方面，NYS染色的测量（必然在福尔根程序之后应用）似乎易受福尔根水解持续时间的影响。这种易感性可能是由于DNA磷酸盐与碱性氨基酸残基的相互作用，NYS的潜在结合位点。由于这种相互作用的程度可能是可变的，有人认为NYS结合将测量染色时蛋白质的可用碱度，但不是特定的蛋白质部分。DNFB结合不受DNA - 蛋白质相互作用的影响，因此可以更可靠地测量“核”蛋白质，特别是与福尔根 - DNA测量结合使用时。
