BOSTER BIOLOGICAL TECHNOLOGY Co.,Ltd.
3942 B Valley Ave, Pleasanton, CA, 94566
Phone: 888-466-3604 Fax: 925-215-2184 Email: [email protected] Web: www.bosterbio.com
RIPA Lysis Buffer
Catalog No.
AR0105
Form Supplied
Ready-to-use 1X solution
Physical State
Liquid
Pack Size
50 ml
Recommended working concentration
10 ml RIPA Lysis Buffer per gram of tissue
1 ml RIPA Lysis Buffer per 2.0x107 cells in suspension
1 ml RIPA Lysis Buffer per 2.0x107 adherent mammalian cells
Compatibility with reagents
Fully compatible with Broad Spectrum Protease Inhibitor Cocktail and
Storage
Broad Spectrum Phosphatase Inhibitor Cocktail
Store at 4˚C for one year.
Description
RIPA Lysis Buffer is a complete cell lysis solution reagent used for rapid and efficient total cell lysis and solubilization of
proteins from both adherent and suspension cultured mammalian cells, effectively extracting cytoplasmic, nuclear and
membrane proteins. RIPA lysis buffer is highly compatible with various downstream protein analysis applications.
Application
Western blotting, ELISA, immunofluorescence, Immunoprecipitation; protein purification; SDS-PAGE;
assays for protein expression, activity, modification, profiling, characterization and quantitative
measurement; epitope tagging; reporter gene analysis
Reagent Type
Western Blotting related reagent; Cell lysis buffer; Universal tissue extraction buffer; Detergent solution
Content
1% NP-40, 0.5% Sodium deoxycholate, 0.1% SDS
Usage
Extraction of cytoplasmic, membrane and nuclear proteins
Protocol
1. Cell sample
FOR RESEARCH USE ONLY. NOT FOR DIAGNOSTIC AND CLINICAL USE.
Product Information Sheet
Adherent cell: Remove culture medium, wash sample with pre-cooling PBS, and scrape cells with scraper (Do not
trypsinize cells to avoid protein degradation). Centrifuge at ~3000 x g for 5 minutes. Remove supernatant and add
precooling RIPA lysis buffer into the cell sample. Incubate on ice for 1-3 hours. And then centrifuge at 10000-14000g for
10 min. Collect the supernatants for the following use.
Suspending cell: Centrifuge at ~3000 x g for 5 minutes to collect cells. Wash sample with pre-cooling PBS. Centrifuge
at ~3000 x g for 5 minutes. Remove supernatant and add precooling RIPA lysis buffer into the cell sample. Incubate on
ice for 1-3 hours. And then centrifuge at 10000-14000g for 10 min. Collect the supernatants for the following use.
Instruction for RIPA USAGE
SIZE of well/surface area
Kit volume
100mm
500-1000μl
60mm
250-500μl
6-well plate
200-400μl per well
24-well plate
100-200μl per well
96-well plate
50-100μl per well
2. Tissue Sample:
Mince tissue and then rinse tissue in ice-cold PBS. And place minced tissue in tissue homogenizer.
Add ice-cold lysis buffer. For a 0.5g piece of tissue, add 5ml of ice-cold lysis buffer (Volumes of lysis buffer must be
determined in relation to the amount of tissue present.)
Homogenize for several minutes at high speed until no tissue chunks remain.
Incubate on ice for 1-3 hours.
Centrifuge at ~10000 x g for 10 minutes. And then aspirate supernatant. The centrifugation force and time can be varied
depending on the sample type.
Note: Precool at 4˚C and add protease inhibitor in RIPA Lysis Buffer several minutes before use.
FOR RESEARCH USE ONLY. NOT FOR DIAGNOSTIC AND CLINICAL USE.
Product Information Sheet