| Cat.No.: | EL-0308 |
| Product Name: | Hep2 Whole Cell Lysate |
| Species: | Human |
| Source: | HEp2 (HeLa contaminant) |
| Preparation Method: | The cells were grown in MEM supplemented with 10% FBS (Fetal Bovine Serum). The lysate was prepared by first washing the cells in PBS supplemented with EDTA and a cocktail of protease inhibitors (see below). Washed cells are then incubated on ice in modified RIPA buffer containing 150 mM sodium chloride, 50 mM Tris Cl, pH 7.4, 1 mM EDTA, 1.0% NP-40, 0.25% sodium deoxycholic acid to lyse the cells. Protein integrity is ensured using a cocktail of protease inhibitors with broad specificity for the inhibition of aspartic, cysteine, and serine proteases as well as aminopeptidases (0.5 mM AEBSF HCl, 0.4 mM Aprotinin, 25 M Bestatin, 7.5 M E-64, 10 M Leupeptin Hemisulfate and 5 M Pepstatin A). Cell debris was removed by centrifugation. Protein concentration was determined by biuret assay using a commercially available kit. The protein concentration was adjusted to 2 mg/ml and then an equal volume of 2X SDS-PAGE sample buffer was added. |
| Recommended Usage: | Protein Lysate for WB. Ready-to-use lysates are especially prepared as positive controls for separation by SDS-PAGE and subsequent Western blotting analysis. Lysates are prepared in denaturing buffer WITHOUT dissociating agents (i.e. no 2- mercaptoethanol or dithiothreitol has been added). |
| Storage Buffer: | 1X SDS-PAGE Sample Buffer (62.5 mM Tris HCl, 2% SDS, 10% Glycerol and 0.005% bromophenol blue, pH 6.8) |
| Storage: | 1X SDS-PAGE Sample Buffer (62.5 mM Tris HCl, 2% SDS, 10% Glycerol and 0.005% bromophenol blue, pH 6.8) |
| Warning: | For research use only, not for diagnostic or therapeutic use. |
| Product Types | ||
| ◆ Extracts & Lysates | ||
| EL-0001 | Recombinant Human MBD1 Cell Lysate | Inquiry |
| EL-0002 | Recombinant Human MBD3 Cell Lysate | Inquiry |
| EL-0003 | Recombinant Human MBD3L1 293 Cell Lysate | Inquiry |
| EL-0004 | Recombinant Human MBD4 Lysate | Inquiry |
| EL-0005 | Recombinant Human MBD5 Cell Lysate | Inquiry |
USA
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