Immunocytochemistry Protocols.
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Microwave protocol: 18°C; Power level 2; 2 min on/2 min off/2 min on/2 min off. Pella research-grade oven (# 3450, with variable power controller #3430 and cold spot). A: sodium azide. Alexa 564-Donkey anti-goat tertiary (Molecular Probes #A11056). BSA: bovine serum albumin, IgG-free (Jackson Immunoresearch #001-000-161). Cy3-Goat anti-mouse secondary (Jackson #115-165-068). Cy3-Goat anti-rabbit secondary (Jackson # 111-165-144). Cy3-Goat anti-rabbit F(ab)2 (Jackson # 111-166-047). DNAse I (Sigma): 10 U DNAse, 4 mM MgCl2 in PBS; 1 hr, 37°C. G: glutaraldehyde. M/F: methanol/37% formalin. NaBH4: 0.01 M NaBH4, 0.5% SMB in 0.05 M Tris HCl, pH 7.5; 30 min. NGS: normal goal serum. ON: overnight on a shaker. P: paraformaldehyde. PB: phospate buffer pH 7.4. PBS: phosphate-buffered saline (10 mM sodium phosphate, 150 mM NaCl, pH 7.4). RT: room temperature. SMB: sodium metabisulfite. T: Triton X-100. TB: 10 mM Tris-HCl, pH 7.4. TBS: Tris-buffered saline (20 mM Tris-HCl, 150 mM NaCl, pH 7.4).
