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U-937 Cell Nuclear Extract
W09-001-GL2
200 µg
Liquid (sterile filtered)
Human
Shipping info:
$50.00 to US & $70.00 to Canada for most products. Final costs are calculated at checkout.
Product Details
U-937 Cell Nuclear Extract - W09-001-GL2
U-937 lysate nuclear extract, cell lysate, U937 Nuclear lysate
Human
Target Details
U-937 cells were grown in RPMI-1640 medium supplemented with 10% fetal bovine serum. 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 4 mg/ml in RIPA buffer containing protease and phosphatase inhibitors.
Application Details
Multi-purpose U-937 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.
Cell Line Data
Human - Pleural Effusion, Lymphocyte, Myeloid
Nuclear Extract
Not Stimulated
Tissue Culture
None (Control)
Formulation
4.0 mg/mL by BCA assay
1X RIPA Buffer with HALT Protease and Phosphatase Inhibitors
None
None
Shipping & Handling
Dry Ice
Store vial at -70° C or COLDER. For extended storage, aliquot Nuclear Extract to minimize freeze/thaw cycles.
Expiration date is three (3) months from date of receipt.
Multi-purpose U-937 nuclear extracts 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.
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.
