GUS histochemical staining Stock:

50mM K3Fe(CN)6                                   0.82315 g/50 ml

50mM K4Fe(CN)6                                   1.056 g/50 ml

1M Na2HPO4                                          7.1 g/50 ml

1M KH2PO4                                     6.8 g/50 ml

0.5M Na2EDTA                                18.612 g/ 100ml

                (直接加入 NaOH crystal，調pH = 8.0 )

10﹪Triton X-100                               1ml/10ml

x-gluc solution(1ml): 50mg/1ml DMSO or DMF

方法：

1.     染色材料在90％的丙酮中固定15－20min，冰上操作（丙酮使蛋白变性）(丙酮，-20预冷)

2.     染色Buffer漂洗后，加入GUS染液（Buffer : x-gluc=1ml :20ul），抽真空5min，37度保温遮光过夜

3.    染色后用酒精梯度脱色(30, 50, 70, 95, 100, and 100%)，体视显微镜观察，拍照。

GUS染色液Buffer:

Stock                                final conc.                              分子量           1L

1M Na2HPO4                100mM           0.64ml     358g/M          22.912g

1M KH2PO4                                          0.36ml     136.09g/M        4.89924g

0.5M Na2EDTA              10mM                      0.2ml       372.2 g/M       3.722g

50mM K3Fe(CN)6         0.5mM                      0.1ml       329.25 g/M     0.16462g

50mM K4Fe(CN)6         0.5mM                      0.1ml       422.39g/M      0.21119g

10﹪Triton X-100           0.1﹪                         0.1ml                      1ml

MeOH（甲醇，有毒）       10﹪                        1ml                      100ml

x-gluc                               0.3﹪         30mg以250μl DMSO