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Thermo Scientific™ RIPA Lysis and Extraction Buffer

Catalog No. PI89900
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250 mL
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PI89901 250 mL
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Catalog No. PI89900 Supplier Thermo Scientific™ Supplier No. 89900
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Description

Lyse cultured mammalian cells with this high-quality, ready-to-use and fully disclosed formulation of a popular cell lysis reagent.

Thermo Scientific RIPA Lysis and Extraction Buffer is a high-quality, ready-to-use and fully disclosed formulation of a popular cell lysis reagent for cultured mammalian cells.

Features of RIPA Buffer:

  • Convenient—ready-to-use solution; no need to assemble and prepared components yourself
  • Flexible—compatible with many applications, including reporter assays, protein assays, immunoassays and protein purification
  • Versatile—enables extraction of cytoplasmic, membrane and nuclear proteins
  • Disclosed formulation—contains no proprietary components, providing users with complete control and knowledge of possible compatibility issues

This RIPA buffer effectively lyses and extracts protein from cultured mammalian cells, including plated cells and pelleted suspension cells. The popular reagent enables the extraction of membrane, nuclear and cytoplasmic proteins and is compatible with many applications, including reporter assays, the Thermo Scientific BCA Protein Assay (Cat. No. 23225), immunoassays and protein purification. Inhibitors such as Thermo Scientific Halt Protease Inhibitor Cocktail (Cat. No. 78420) and Halt Phosphatase Inhibitor Cocktail (Cat. No. 78420) are also compatible with this RIPA buffer formulation and can be added before use to prevent proteolysis and maintain protein phosphorylation.

RIPA buffer derives its name from the original application for which it was developed: the radio-immunoprecipitation assay. While this isotopic assay method is rarely performed in laboratories today, the acronym for this lysis buffer formulation has endured in common use. RIPA cell lysis reagent is highly effective for protein extraction from a variety of cell types because it contains three non-ionic and ionic detergents. One disadvantage of this detergent formulation is its relative incompatibility with certain downstream applications compared to other lysis reagents.

Related Products

  • Pierce™ IP Lysis Buffer (Cat. No. 87787)
  • M-PER™ Mammalian Protein Extraction Reagent (Cat. No. 78501)

Specifications

Quantity 100 mL
Format Liquid
Product Type Extraction Buffer
Volume (Metric) 100 mL
Content And Storage Upon receipt store at 4°C.

Frequently Asked Questions (FAQs)

What are the standard lysis buffers used with mammalian cells for detection of protein expression by immunoprecipitation (IP) or Western blot analysis?

The most commonly used buffer is RIPA Buffer with SDS. We offer RIPA Buffer (Cat. Nos. 89900 and 89901). We also offer the Pierce IP Lysis buffer (Cat. Nos. 87787 and 87788) as well as M-PER (Cat. Nos. 78501, 78503, and 78505).
Does RIPA Lysis and Extraction Buffer (Cat. No. 89900, 89901) contain protease or phosphatase inhibitors?

RIPA Lysis and Extraction Buffer (Cat. No. 89900, 89901) does not contain protease or phosphatase inhibitors. If desired, you may add protease and/or phosphatase inhibitors, such as Halt Protease Inhibitor Cocktail (Cat. No. 78410) and Halt Phosphatase Inhibitor Cocktail (Cat. No. 78420) to the RIPA Lysis and Extraction Buffer to prevent proteolysis and maintain phosphorylation status of proteins. We recommend adding protease and phosphatase inhibitors immediately before use.
Why is it not recommended that I use RIPA buffer for protein A280 measurements with my NanoDrop spectrophotometer?

RIPA buffer produces a particularly strong absorbance signal at the 280 nm wavelength. As a result, it will either over estimate or under estimate protein concentrations and interfere with the protein purity ratio.
Protein samples in RIPA buffer should be quantified via the Pierce Protein 660 or BCA colorimetric assays using a full spectrum NanoDrop model.

Why is there low phosphorylation of the proteins when I use the RIPA Lysis and Extraction Buffer?

Low phosphorylation is usually due to phosphatase activity. We recommend adding a Halt Phosphatase Inhibitor Cocktail to the buffer before use.
Alternatively, the protein is not phosphorylated or phosphorylated at a low level.
What if proteolysis occurs when I use RIPA Lysis and Extraction Buffer?

Proteolysis indicates that no protease inhibitors were added. We recommend adding a Halt Protease Inhibitor Cocktail to the RIPA Lysis and Extraction Buffer before use.

I am getting a low concentration of proteins when using the RIPA Lysis and Extraction Buffer. What could be a possible cause and solution?

Most likely excess buffer was used. Use less buffer (e.g., 0.25 mL to 0.5 mL per 75cm^2 flask containing 5 x 10^6 cells), but use a sufficient amount to cover the entire plate.

Why is the total protein yield low when I use the RIPA Lysis and Extraction Buffer?

Some cells are more resistant to lysis than others and can cause a low protein yield. Make sure the cell pellet is thoroughly suspended in RIPA Buffer, incubate for longer with occasional swirling, and sonicate the pellet to increase yield.

For Research Use Only. Not for use in diagnostic procedures.