Radioimmunoprecipitation assay buffer is a lysis buffer

Lysis buffer

A lysis buffer is a buffer solution used for the purpose of lysing cells for use in molecular biology experiments that analyze the compounds of the cells . There are many different kind of lysis buffer that one can apply, depending on what analysis the cell lysate will be used for e.g. red blood...

 used to lyse

Lysis

Lysis refers to the breaking down of a cell, often by viral, enzymic, or osmotic mechanisms that compromise its integrity. A fluid containing the contents of lysed cells is called a "lysate"....

 cells and tissue, for radio immunoprecipitation

Immunoprecipitation

Immunoprecipitation is the technique of precipitating a protein antigen out of solution using an antibody that specifically binds to that particular protein. This process can be used to isolate and concentrate a particular protein from a sample containing many thousands of different proteins...

 assay

Assay

An assay is a procedure in molecular biology for testing or measuring the activity of a drug or biochemical in an organism or organic sample. A quantitative assay may also measure the amount of a substance in a sample. Bioassays and immunoassays are among the many varieties of specialized...

 (RIPA). This buffer is more denaturing than NP-40 or Triton X-100 lysis buffer because it contains the ionic detergents SDS and sodium deoxycholate as active constituents and is particularly useful for nuclear membrane disruption for nuclear extracts. The RIPA buffer gives low background but can denature kinases.

Recipe

RIPA buffer recipe for total protein

Protein

Proteins are biochemical compounds consisting of one or more polypeptides typically folded into a globular or fibrous form, facilitating a biological function. A polypeptide is a single linear polymer chain of amino acids bonded together by peptide bonds between the carboxyl and amino groups of...

 extract is as follows:

• Tris 50mM
• NaCl 150 mM
• SDS 0.1 %
• Na.Deoxycholate 0.5 %
• Triton X 100 or NP40 1%
• Protease
  Protease
  A protease is any enzyme that conducts proteolysis, that is, begins protein catabolism by hydrolysis of the peptide bonds that link amino acids together in the polypeptide chain forming the protein....
  
  inhibitors such as PMSF or cocktail protease inhibitors which are commercially available (use according to the manufacturer's guide)

External links

• 1X RIPA Buffer