Selected article for: "sample buffer and SDS PAGE sample buffer"
Title: Constitutive and basal secretion from the endocrine cell line, AtT-20
  • Document date: 1991_3_1
    • ID: tqgsavnr_13
    Snippet: Sulfated proteins were labeled by incubating cells for 18 h at normal growth conditions of 37°C and 10-15% CO2, in sulfate-depleted DME H21 conraining 5 mCi [35S]sulfate, penicillin, streptomycin, glutamine, and 10% dialyzed FCS as described (Moore et al., 1983a) . Cells were then incubated in serum-free DME H21 for two 1.5-h intervals, the latter interval in the presence or absence of 5 mM 8-Br-cAMP. Samples were precipitated with cold 10% TCA .....
    Document: Sulfated proteins were labeled by incubating cells for 18 h at normal growth conditions of 37°C and 10-15% CO2, in sulfate-depleted DME H21 conraining 5 mCi [35S]sulfate, penicillin, streptomycin, glutamine, and 10% dialyzed FCS as described (Moore et al., 1983a) . Cells were then incubated in serum-free DME H21 for two 1.5-h intervals, the latter interval in the presence or absence of 5 mM 8-Br-cAMP. Samples were precipitated with cold 10% TCA containing 1 mg/ml deoxycholate, washed twice with cold acetone, and redissolved in SDS-PAGE sample buffer. Analysis was performed on 12-20% polyacrylamide gradient gels, which were soaked in 0.5 M sodium salicylate containing 30% methanol, dried, and exposed to film (XAR-5; Eastman Kodak Co., Rochester, NY).

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