| 产品详情 |
| Edit |   |
|---|
| Product Name | MCF-7 H2O2 Stimulated Whole Cell Lysate |
|---|
| Description | The cells were grown in Dulbecco'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 2 mg/ml and then an equal volume of 2X SDS-PAGE sample buffer was added. |
|---|
| Size | 500 µg |
|---|
| Concentration | 1.0 mg/mL |
|---|
| Applications | 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). Heat lysate to 95° C for 5 minutes and rapidly cool. If dissociating conditions are desired, add reducing agent prior to heating. The recommended loading volume per lane is 10-20 µl depending on the size format of your gel. |
|---|
| Other Names | MCF-7 H2O2 Stimulated Whole Cell Lysate, MCF7 Hydrogen Peroxide Stimulated WCL, MCF-7 Peroxide Lysate |
|---|
| Gene, Accession, CAS # | n/a |
|---|
| Catalog # | W09-000-366 |
|---|
| Price | |
|---|
| Order / More Info | MCF-7 H2O2 Stimulated Whole Cell Lysate from ROCKLAND IMMUNOCHEMICALS INC. |
|---|
| Product Specific References | n/a |
|---|
| 产品资料 |
|
|