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HeLa Whole Cell Lysate MG-132 Treated
W09-001-GV9
500 µg
Liquid
SDS-PAGE
Human
Shipping info:
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Product Details
HeLa Whole Cell Lysate - MG-132 Stimulated - W09-001-GV9
HeLa Lysate MG132 stimulated, Cell Lysate, MG-132 Stimulated Lysate HeLa cell, HeLa lysate MG-132 treated
Human
Target Details
HeLa cells were grown in Dulbecco’s medium supplemented with 10% fetal bovine serum. Cells were treated O/N with 0.2µg/mL of MG-132 in order to reduce degradation of ubiquitin-conjugated proteins by the proteasome. Cells were washed with PBS and then incubated on ice in modified RIPA buffer 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.
Application Details
SDS-PAGE
W09-001-GV9 has been tested by SDS-PAGE. 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.
Cell Line Data
HeLa - Human epidermoid carcinoma
Whole Cell Lysate
MG-132
Tissue Culture
MG-132
Formulation
1mg/mL by modified Lowry assay
1X SDS-PAGE Sample Buffer (62.5 mM Tris HCl, 2% SDS, 10% Glycerol and 0.005% bromophenol blue, pH 6.8)
None
10% (v/v) Glycerol
Shipping & Handling
Dry Ice
Store vial at -70° C or COLDER. For extended storage, aliquot contents to minimize freeze/thaw cycles.
Expiration date is three (3) months from date of receipt.
Ready-to-use whole cell lysates produced by Rockland Immunochemicals are derived from cell lines or tissues using highly refined extraction protocols to ensure exceptionally high quality, protein integrity and lot-to-lot reproducibility. All extracts are tested by SDS-PAGE using 4-20% gradient gels and immunoblot analysis using antibodies to key cell signaling components to confirm the presence of both high molecular weight and low molecular weight proteins.
This product is for research use only and is not intended for therapeutic or diagnostic applications. Please contact a technical service representative for more information. All products of animal origin manufactured by Rockland Immunochemicals are derived from starting materials of North American origin. Collection was performed in United States Department of Agriculture (USDA) inspected facilities and all materials have been inspected and certified to be free of disease and suitable for exportation. All properties listed are typical characteristics and are not specifications. All suggestions and data are offered in good faith but without guarantee as conditions and methods of use of our products are beyond our control. All claims must be made within 30 days following the date of delivery. The prospective user must determine the suitability of our materials before adopting them on a commercial scale. Suggested uses of our products are not recommendations to use our products in violation of any patent or as a license under any patent of Rockland Immunochemicals, Inc. If you require a commercial license to use this material and do not have one, then return this material, unopened to: Rockland Inc., P.O. BOX 5199, Limerick, Pennsylvania, USA.
