MDA-MD-435S Whole Cell Lysate
Cat.No. : | MDA-MD-435S-20H |
- Specification
- Gene Information
- Related Products
Species : | Human |
Concentration : | 2 mg/ml |
Tissue Type : | mammary gland ductal carcinoma |
Preparation method : | The cells were grown in Leibovitz's L-15 medium with 0.01 mg/ml insulin, 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 and 0.1% SDS, 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 M Aprotinin, 5 M Bestatin, 1.5 M E-64, 2 M Leupeptin Hemisulfate, 1 M 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 2 mg/ml and then an equal volume of 2X SDS-PAGE sample buffer was added. |
Recommended Usage : | For research use only, not for diagnostic or therapeutic use. |
Storage Buffer : | 1X SDS-PAGE Sample Buffer (62.5 mM Tris HCl, 2% SDS, 10% Glycerol and 0.005% bromophenol blue, pH 6.8) |
Applications : | Protein Lysate for WB. Ready-to-use lysates are especially prepared as positive controls for separation by SDS-PAGE and subsequent western blot analysis. Lysates are prepared in denaturing buffer WITHOUT dissociating agents (i.e. no 2- mercaptoethanol or dithiothreitol has been added). |
For Research Use Only. Not intended for any clinical use. No products from Creative BioMart may be resold, modified for resale or used to manufacture commercial products without prior written approval from Creative BioMart.
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