Cells were serum starved for 48 h, then seeded to a pretreated (incubated 20 min in Radius Gel Pretreatment solution and washed with Radius Wash Solution) Radius 96-Well Plate at a density of 8 × 104 cells per well with or without E2 (10 nM). After 24 h incubation, the ...
Standard sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and Western blotting procedures were used to analyse the expression of various proteins. Cell lysates were prepared using SDS lysis buffer (50 mM Tris-HCl, pH 6.8, 2% SDS, 10% glycerol, and 0.02% bromophenol blue) ...
Samples were loaded onto an Any kD Mini-PROTEAN TGX Precast Protein Gel (Bio-Rad) along with 5 μl of a protein ladder (Bio-Rad). After the gel electrophoresis, the transfer onto a methanol-activated PVDF membrane (Bio-Rad) was achieved by wet blotting for 1 h at 350 mA. Membranes ...
Proteins were resolved by SDS–PAGE (polyacrylamide gel electrophoresis) and transferred to AmershamHybond-P PVDF membranes (GE Healthcare), followed by blocking with 5% non-fat dry milk. The following antibodies were purchased from Cell Signaling Technology, Inc. and used at a 1:1,000 dilution...
Cells were harvested in RIPA-lite lysis and resolved using sodium dodecyl sulfate-polyacrylamide gel electrophoresis as previously described.15 Immunoblotting was performed with the following antibodies: PR (#sc-166169; Santa Cruz Biotechnology), pS294 PR (made in-house), IGF1Rβ (#3027 S; Cel...
LacZactivity was measured in a semiquantitative X-Gal overlay assay. The interacting ‘prey’ fragments of the positive clones were amplified by PCR, analysed on an agarose gel, and sequenced at their 5′ and 3′ junctions on a PE3700 Sequencer. The resulting sequences were then used to ...
Prat A, Cheang MC, Martin M, Parker JS, Carrasco E, Caballero R, Tyldesley S, Gelmon K, Bernard PS, Nielsen TO, Perou CM (2013) Prognostic significance of progesterone receptor-positive tumor cells within immunohistochemically defined luminal A breast cancer. J Clin Oncol 31: 203–209. ...
(cell lines with and without ERα or ERβ) were analyzed by MS. Information on each identified peptide is provided, along with gene name, protein name, expected MW, molecular function and corresponding transcript level (RNA-seq of corresponding cell lines) for each separate gel bands analyzed....