Phosphate-buffered saline (PBS): 8 g NaCl, 0.2 g KCl, 1.44 g Na2HPO4 and 0.24 g KH2PO4 per litre at pH 7.2
Halogenated nucleosides: stock solutions of 10 mg/ml of 5-bromo-2'-deoxyuri-dine (BrdU, Sigma), 5-chloro-2'-deoxy-uridine (CldU, Sigma), or 5-iodo-2'-deoxyuridine (IdU, Sigma) are prepared in cell culture medium (without serum) at pH 7.2. Store frozen at -20 °C in small aliquots
Antibodies: (A) Mouse anti-BrdU (No. 347580; Becton Dickinson) is used for detection of BrdU, CldU and IdU . (B) Rat anti-BrdU (MAS250b; HarlanSera lab) is used for detection of BrdU and CldU . (C) Secondary antibodies: FITC-conjugated donkey anti-rat IgG (No. 712-095-153; Jackson ImmunoRe-search Laboratories) or FITC-conjugated donkey anti-mouse IgG (No. 715-095151; Jackson ImmunoResearch Laboratories) are used to detect the rat or mouse anti-BrdU antibodies, respectively
Blocking buffer: to suppress non-specific binding of antibodies to coverslips or to cellular material, antibody dilutions are made in PBS containing 2% bovine serum albumin and either 10% normal donkey serum, or 10% fetal bovine serum. Supplement with NaN3 at 1 mM final concentration and store at 4 °C
DNA staining dye: a stock solution of 1 mg/ml of 4',6-diamidino-2-phenylin-dole (DAPI; Sigma) is prepared in ddH2O and stored frozen in small aliquots at -20 °C. A working solution of 10 ^g/ml can be kept at 4 °C
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