Sterile; Ultrapure, Filtered 0.1-micron membrane TWICE and 0.04-micron membrane ONCE.
World’s cleanest buffer for all cell and molecular biology experiments
Description:
RIPA (Radioimmunoprecipitation Assay) buffer is designed to extract cytoplasmic, membrane, and nuclear proteins. It is a strong ionic detergent-based lysis buffer that disrupts cell and nuclear membranes to release proteins. It contains sodium deoxycholate which is useful for nuclear membrane disruption for nuclear extracts. It can be used to lyse both adherent and suspension-cultured cells. The only difference is when introducing the RIPA buffer to the cell culture. For adherent cells, add the RIPA buffer directly to the washed cell monolayer after washing the cells with a wash buffer and use a cell scraper to collect the lysate. This scraping can be done over ice to maintain the cold temperature during lysis. For suspension cells, the cell pellet is resuspended in the RIPA buffer after washing cells.
* The RIPA buffer does not contain protease or phosphatase inhibitors, so these should be added if needed to prevent protein degradation and maintain phosphorylation states.
Advantages:
It can extract a wide range of proteins.
The combination of ionic (SDS, sodium deoxycholate) and non-ionic (NP-40) detergents in RIPA buffer effectively disrupts cell membranes and solubilizes proteins, making it effective for extracting proteins that are otherwise difficult to solubilize.
Compatibility with protease and phosphatase inhibitors.
The components of the RIPA buffer do not interfere with common protein quantification methods like the BCA assay.
Applications:
The strong detergents in RIPA buffer, such as SDS and sodium deoxycholate, help denature proteins and disrupt protein-protein interactions, making them suitable for Western blotting.
RIPA buffer is compatible with immunoprecipitation experiments, as it can disrupt non-specific protein-protein interactions while preserving specific interactions of interest.
The ability of the RIPA buffer to solubilize a wide range of proteins makes it useful for extracting proteins before purification.
RIPA buffer-extracted proteins can be used in reporter assays and protein quantification methods, such as the BCA assay.
Composition:
Ingredients
Concentration
Tris-HCl
50 mM
NaCl
150 mM
Nonidet P-40
1%
Sodium deoxycholate
0.5%
SDS
0.1%
EDTA
1 mM
Quality Control:
Appearance: Clear, Colorless Liquid.
pH: 7.4
Sterility: Filtered 0.1 micron Twice and 0.04 micron once in a sterile environment.
DNase activity: None was detected after incubation of plasmid DNA and this product for 18 hr at room temperature.
RNase activity: No RNase activity was detected after incubation of ribosomal RNA and this product for 18 hr at room temperature.
Storage and Shelf Life:
Storage: 4 ℃
Shelf life: 2 years
CUSTOMIZATION: Please INQUIRE if other concentrations, additions of chemicals/compounds/proteins/supplements, different pH, and modifications are needed.
All DiagnoCine Precision Sterile buffers are filtered-sterilized with a 0.1-micron filtration two times and 0.04-micron filtration one time. Thus, mycoplasma contamination is prevented. The smallest size mycoplasma type can be about 0.2 microns.
* This product is manufactured under ISO 13485-certified and CE-approved facilities (Suppliers of DiagnoCine Precision). All final packaging, quality assurance, and testing are done at the DiagnoCine R&D and Quality Testing Center. All specific customization requests and assembly were accomplished at DiagnoCine Precision in Totowa, New Jersey, USA
