Immunohistochemical technology of labeling skin of inner mongolia cashmere goats with BrdU
- VernacularTitle:BrdU标记山羊皮肤组织的免疫组化检测
- Author:
Haiyan XI
;
Yan ZHENG
;
Huanmin ZHOU
- Publication Type:Journal Article
- Keywords:
5-bromo-2-deoxyuridine;
Cashmere goat;
Immunohistochemistry;
Labeling
- From:
Chinese Journal of Immunology
2001;0(07):-
- CountryChina
- Language:Chinese
-
Abstract:
Objective:The purpose of this study is to investigate the feasibility of labeling skin of inner mongolia cashmere goats with BrdU,and to optimize its immunohistochemical conditions.Methods:We injected different dose of BrdU into the neck vein of cashmere goats,then we took samples from the back skin of the goats,fixed in 4% paraformaldehyde.Next the samples were embedded with paraffin and sectioned.We examined the samples with immunohistochemical technology to test the effect of the influencing factors to labeling results as the BrdU injection,such as dosage,the concentration of second antibody and streptavidin-peroxidase,the way of antigen pretreatment,and DNA denature temperature.Results:We observed red BrdU positive cells in labeled goats whereas not in the control.Meanwhile,in the samples of the unlabeled goats or those incubated with PBS,no nuclear staining is observed.Conclusion:BrdU can be used for labeling of inner mongolia cashmere goats in vivo,which have no obvious dosage effect.The best condition of labeling is that puttig the section in 2 mol/L HCl 30 min at 37℃,in trypsin 30 min at 37℃.The most appropriate concentration of biotin-conjugated second antibody and streptavidin-peroxidase is 1∶100,under which the results of immunohistochemical are the best.
