The Thermo Scientific Pierce IP Lysis Buffer is optimized for cell lysate yield, purity and compatibility with immunoprecipitation (IP and Co-IP) as the downstream application for the cell lysate.
The IP Lysis Buffer is a mammalian whole cell lysis reagent based on a modified RIPA buffer formulation without SDS. This moderate-strength lysis buffer effectively solubilizes cellular proteins but does not liberate genomic DNA or disrupt protein complexes like ordinary RIPA buffer. Pierce IP Lysis Buffer is specially formulated for pull-down and immunoprecipitation assays.
- Optimized for compatibility with immunoprecipitation and pull-down assays
- Compatible with protein assays, reporter assays and immunoassay procedures
- Ready-made formula is effective for extracting cytoplasmic, membrane and nuclear proteins
- Gentle formulation helps maintain protein complexes for co-immunoprecipitation
- Does not liberate genomic DNA which can cause high sample viscosity
The Pierce IP Lysis Buffer is effective for lysing cultured mammalian cells from both plated cells and cells pelleted from suspension cultures. Optimized for pull-down and immunoprecipitation assays, this lysis buffer is also compatible with many other applications, including the Thermo Scientific Pierce BCA and 660 nm Protein Assays, protein purification and immunoassays (e.g., ELISA, Western blot).
The Pierce IP Lysis Buffer is composed of 25mM Tris-HCl pH 7.4, 150mM NaCl, 1mM EDTA, 1% NP-40 and 5% glycerol. The buffer does not contain protease or phosphatase inhibitors; however, if desired, inhibitors, such as Thermo Scientific Halt Protease Inhibitor Cocktail or Phosphatase Inhibitor Cocktail can be added just before use to prevent proteolysis and maintain phosphorylation of proteins.
|The moderate-strength Pierce IP Lysis Buffer produces high yield. A549 cells (2 x 10^6) were lysed using 200µL of three different lysis buffers, clarified by centrifugation and measured for protein concentration using the Thermo Scientific Pierce BCA Protein Assay (Part No. 23227).
|Pierce IP Lysis Buffer enables highly efficient immunoprecipitations. A549 cell lysates (200µg) were generated using different lysis buffers. EGFR and PP2A were immunoprecipitated using the Thermo Scientific Pierce Crosslink IP Kit (Part No. 26147) and anti-EGFR antibody (Stressgen) or anti-PP2A antibody (Millipore). The immunoprecipitations were washed with the same lysis buffer used to lyse the cells. Lysates representing 5% of the total load (L) and elution (E) fractions were blotted with anti-EFGR or anti-PP2A antibody and using Thermo Scientific SuperSignal West Dura Substrate (Part No. 34076) for detection.
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Immunoprecipitation and Pull-Down Kits