Non-Abcam antibody was used: CY3 GOAT ANTI MOUSE IgG Host species: Goat Clonality: Polyclonal Conjugation: Cy3 ®
Slides were rehydrated and then boiled for antigen retrieval in Citrate pH 6 buffer for 15min in standard microwave oven at 650W. After 30min blocking in PBS-Triton 0,1% / NGS 10% slides were incubated O/N with primary mAbcam 51841 (TBP) antibody at 1/400 dilution and ab15102 (Claudin3) antibody at 1/300 dilution at 4°C. The secondary antibody was CY3 GOAT ANTI MOUSE IgG (for TBP) and Alexa Fluor® 488 goat anti-rabbit IgG (H+L) (for Claudin3) both used at a 1/200 dilution for 1h. Nuclei were stained with Hoechst. Slides were mounted with VECTASHIELD mounting solution.
Submitted May 14 2013
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