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目的 :探讨H1 2 单克隆抗体的不同浓度对FITC荧光标记结果的影响。方法 :紫外吸收法测定抗体蛋白质浓度 ,透析标记法进行抗体荧光标记 ,并以ELISA法对抗体进行特异性鉴定及效价分析。结果 :标记前后抗体特异性经ELISA鉴定未见明显差异。结论 :①FITC荧光标记前抗体蛋白质浓度应达 (10～ 2 0 )mg mL ,才能得到合适的F P比值。②F P值在 1～ 2之间者较为理想 ,如果过标记的抗体F P >2则可导致非特异性染色的增加 ,增加假阳性率 ;标记不足F P <1则可导致假阴性率增加。③标记后虽可出现特异性下降和蛋白量的丢失 ;但特异性鉴定与标记前未见明显差异。故标记后抗体H1 2 的特异性适用于肿瘤组织与正常脑组织间界限的确定
Objective: To investigate the effect of different concentrations of H1 2 monoclonal antibody on FITC fluorescence labeling results. Methods: Antibody protein concentration was determined by ultraviolet absorption method. Antibody fluorescent labeling was performed by dialysis labeling method. Antibody was identified by ELISA and titer analysis was performed. Results: There was no significant difference in antibody specificity before and after labeling by ELISA. Conclusions: ① The protein concentration of FITC pre-fluorescent antibody should reach (10 ~ 20) mg mL to get the appropriate F P ratio. ②F P value of 1 to 2 between the ideal, if the over-labeled antibody F P> 2 can lead to non-specific staining increased, increasing the false positive rate; lack of labeling F P <1 can lead to increased false negative rate. ③ Although there may be a specific marker decreased and protein loss; However, no significant difference between the specific identification and labeling before. Therefore, the specificity of labeled antibody H1 2 is suitable for determining the boundary between tumor tissue and normal brain tissue