Immunohistochemistry
The expression of PSMA was stained by immunohisto- chemistry. The reagent kits were purchased from Fuzhou Maxim Co. Ltd. PSMA monoclonal antibodies were pur- chased from Abcam (Ab19071) (1:350) and Santa Cruz Co. Ltd (sc-69665) (1:100). Homogenous animal IgG that con- tained the same protein quantity was used instead of the first antibody in negative control. The specimens, fixed with 10 % neutralformaldehydesolutionandembeddedinparaffin,were cut into 5-lm-thick sections. Tissue sections were deparaffi- nized and rehydrated with xylene and graded alcohols. Anti- gen retrieval was carried out in 5 mM citrate buffer. After inactivation of endogenous peroxidase with 3 % H2O2 for 10 min, the sections were blocked with goat serum and incubatedwithprimary antibodysolutionat4 C overnightin a humidified chamber. After washing three times with PBS, thesectionsincubatedfor30 minwith biotinylatedsecondary antibody (polyperoxidase-anti-mouse/rabbit IgG, Maixin Corporation, Fuzhou, China) and then washed again with PBS. Diaminobenzidine was used as a chromogen substrate. Sections were then counterstained with hematoxylin for 1 min, raised in water, dehydrated in ascending concentra- tions of ethanol followed by clearance with xylene and cover slipped permanently for light microscopy.
ImmunohistochemistryThe expression of PSMA was stained by immunohisto- chemistry. The reagent kits were purchased from Fuzhou Maxim Co. Ltd. PSMA monoclonal antibodies were pur- chased from Abcam (Ab19071) (1:350) and Santa Cruz Co. Ltd (sc-69665) (1:100). Homogenous animal IgG that con- tained the same protein quantity was used instead of the first antibody in negative control. The specimens, fixed with 10 % neutralformaldehydesolutionandembeddedinparaffin,were cut into 5-lm-thick sections. Tissue sections were deparaffi- nized and rehydrated with xylene and graded alcohols. Anti- gen retrieval was carried out in 5 mM citrate buffer. After inactivation of endogenous peroxidase with 3 % H2O2 for 10 min, the sections were blocked with goat serum and incubatedwithprimary antibodysolutionat4 C overnightin a humidified chamber. After washing three times with PBS, thesectionsincubatedfor30 minwith biotinylatedsecondary antibody (polyperoxidase-anti-mouse/rabbit IgG, Maixin Corporation, Fuzhou, China) and then washed again with PBS. Diaminobenzidine was used as a chromogen substrate. Sections were then counterstained with hematoxylin for 1 min, raised in water, dehydrated in ascending concentra- tions of ethanol followed by clearance with xylene and cover slipped permanently for light microscopy.
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