Application Note: This product is ready-to-use as a working 1X solution and requires no further dilution. 1X RIPA Lysis Buffer is intended for the extraction of cellular proteins for the efficient lysis of cells and solubilization of protein, while minimizing protein degradation and maintaining protein immunoreactivity and biological activity. We recommend using 1.0 ml of RIPA Lysis Buffer to lyse 0.5 to 5 x 10E7 adherent mammalian cells. This buffer contains ionic detergents and may not be suitable for kinase enzymes, if these enzymes are easily denatured. Do not add phosphatase inhibitors when preparing lysates for phosphatase assays. 1X RIPA lysis buffer consists of 50 mM Tris HCl, 150 mM NaCl, 1.0% (v/v) IGEPAL® CA-630, 0.5% (w/v) Sodium Deoxycholate, 1.0 mM EDTA, 0.1% (w/v) SDS and 0.01% (w/v) sodium azide at a pH of 7.4. This buffer was meticulously prepared using ultra pure reagents dissolved in highly polished pharmaceutical grade deionized water.Protease and phosphatase inhibitors are recommended but not included in product composition.Recommended final concentrations of protease inhibitors: 1.0 mM Phenylmethylsulfonyl fluoride (PMSF)10 µM Leupeptin 0.1 µM Aprotinin 1.0 µM PepstatinRecommended final concentrations of phosphatase inhibitors:1.0 mM Na3VO4 1.0 mM NaF
Concentration Value: 1X
General Disclaimer Note: 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.
Physical State: Liquid (sterile filtered)
Purity and Specificity: This product was aseptically filtered through a Millipore 0.22 micron filter into clean, pre-sterilized containers. The product was tested on trypticase soy agar for 24 hours, 48 hours and 72 hours and was found to be negative for bacteria.
Background: RIPA (Radio-Immunoprecipitation Assay) Lysis Buffer enables rapid, efficient cell lysis and solubilization of proteins from both adherent and suspension cultured mammalian cells. It has long been a widely used lysis and wash buffer for small-scale affinity pull-down applications, such as immunoprecipitation, since most antibodies and protein antigens are not adversely affected by the components of this buffer. In addition, RIPA Lysis Buffer minimizes non-specific protein-binding interactions to keep background low, while allowing most specific interactions to occur, enabling studies of relevant protein-protein interactions. The following RIPA Lysis Buffer components have the following effects: Tris-HCl is a buffering agent prevents protein denaturation, NaCl is a salt that prevents non-specific protein aggregation, IGEPAL® CA-630 is a non-ionic detergent to extract proteins; Na-deoxycholate and SDS are ionic detergents to extract proteins; and sodium azide is a bacteriostatic agent added to retard bacterial growth. RIPA Lysis Buffer is supplied as a ready-to-use solution that requires no preparation. We suggest that the user add protease and phosphatase inhibitors not included with this product prior to use.
Low Endotoxin: No
Other: This product is ready-to-use as a working 1X solution and requires no further dilution.
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