产品名称 HL-60 Cell Nuclear Extract from ROCKLAND IMMUNOCHEMICALS INC.
产品货号 W09-001-GK9
产品价格 现货询价,电话:010-67529703
产品规格 200 µg
产品品牌 ROCKLAND IMMUNOCHEMICALS INC.
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Product NameHL-60 Cell Nuclear Extract
DescriptionHL-60 cells were grown in Ioscove's medium supplemented with 10% fetal bovine serum. Cells were washed with PBS and then incubated on ice in modified RIPA buffer, containing 150 mM sodium chloride, 50 mM Tris HCl, pH 7.4, 1 mM EDTA, 1.0% NP-40, 0.5% sodium deoxycholic acid , 0.1% SDS and 0.01% (w/v) sodium azide to lyse the cells. Protein integrity was ensured using a cocktail of protease inhibitors with broad specificity for the inhibition of aspartic, cysteine, and serine proteases as well as aminopeptidases (0.1 mM AEBSF HCl, 0.08 µM Aprotinin, 5 µM Bestatin, 1.5 µM E-64, 2 µM Leupeptin Hemisulfate, 1 µM Pepstatin A). Phosphatase inhibitors 1 mM NaF and 1 mM Na3VO4 were also added. Cell debris was removed by centrifugation. Protein concentration was determined by a modified Lowry assay using a commercially available kit. Protein concentration was adjusted to 4 mg/ml in RIPA buffer containing protease and phosphatase inhibitors.
Size200 µg
Concentration4.0 mg/mL
ApplicationsChip, Immunoprecipitation, Western Blot. Multi-purpose HL-60 nuclear extracts are especially prepared as positive control for multiple assays including western blot, immunoprecipitation (IP), capture ELISA or other assays requiring native protein sample. For separation by SDS-PAGE and subsequent western blot analysis, lysates should be diluted by user to desired concentration in SDS-PAGE buffer with 2-mercaptoethanol or dithiothreitol as the reducing agent and heated to 95° C for 5 minutes. Sample is ready for use in immunoprecipitation and ELISA experiments, conditions should be optimized by the user. Rockland recommends its Trueblot IP reagents for immunoprecipitation experiments.
Other NamesHL-60 lysate nuclear extract, HL60 nuclear lysate, HL-60 Nuclear lysate
Gene, Accession, CAS #n/a
Catalog #W09-001-GK9
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Order / More InfoHL-60 Cell Nuclear Extract from ROCKLAND IMMUNOCHEMICALS INC.
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