Sentences with phrase «coverslips plated»
Coverslips plated with acutely dissociated hNPCctx - GDNF or hNPCctx were fixed with 4 % paraformaldehyde, washed with PBS, blocked in 5 % normal donkey serum and 0.1 % Triton X-100, and incubated with goat anti-GDNF (1 ∶ 100; R&D Systems) primary antibody followed by donkey anti-goat Cy3 - conjugated secondary antibody (1 ∶ 1000; Jackson IR).
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Cells were resuspended in HEPES buffer containing the following (in mm): 136 NaCl, 5.6 KCl, 1 MgCl2, 2.2 CaCl2, 11 glucose, 10 HEPES; pH adjusted to 7.4 with NaOH before plating on poly - d - lysine (0.02 mg / ml; BD Bioscience) and laminin (0.02 mg / ml; Sigma - Aldrich)- coated coverslips.
Cells were plated either on glass coverslips (40,000 cells / coverslip) coated with poly - L lysine and laminin or six - well plates (106 cells / well) coated with laminin alone.
Bulk - cultured PC3 - GFP cells (A), holoclone P1H8 (B)-- , or meroclone P2H3 (C)-- derived cells were plated on glass coverslips and stained for CD44, α2β1, and / or β - catenin using monoclonal antibodies.
Bulk - cultured PC3 or PC3 - GFP cells, or clone - derived cultures were plated on glass coverslips and used in immunofluorescence staining for CD44, α2β1 integrin, and β - catenin using monoclonal antibodies and protocols described previously (27 — 29).