Lysis Buffer
T-Pro RIPA Lysis Buffer (1X)
50 mM Tris HCl, 150 mM NaCl, 1.0% (v/v) NP-40, 1% (w/v) Sodium Deoxycholate and 0.1% (w/v) SDS at a pH of 7.4
T-Pro RIPA Lysis Buffer (5X)
250 mM Tris HCl, 750 mM NaCl, 5.0% (v/v) NP-40, 5% (w/v) Sodium Deoxycholate and 0.5% (w/v) SDS at a pH of 7.4
T-Pro RIPA Lysis Buffer is ready-to-use as a working 1X solution and requires no further dilution. T-Pro RIPA Lysis Buffer is intended for the extraction of cellular proteins for the efficient lysis of cells and solubilization of protein, while minimizing protein degradation and maintaining protein immunoreactivity and biological activity. We recommend using 1.0 mL of RIPA Lysis Buffer to lyse 0.5 to 5 x 10⁷ adherent mammalian cells. This buffer contains ionic detergents and may not be suitable for kinase enzymes, if these enzymes are easily denatured. Do not add phosphatase inhibitors when preparing lysates for phosphatase assays. 1X T-Pro RIPA lysis buffer consists of 50 mM Tris HCl, 150 mM NaCl, 1.0% (v/v) NP-40, 1% (w/v) Sodium Deoxycholate and 0.1% (w/v) SDS at a pH of 7.4.
This buffer was meticulously prepared using ultrapure reagents dissolved in highly polished pharmaceutical grade deionized water. Protease and phosphatase inhibitors are recommended but not included in product composition.
Recommended final concentrations of protease inhibitors:
l 1.0mM Phenylmethylsulfonyl fluoride (PMSF),
l 10 µM Leupeptin,
l 10µM Aprotinin,
l 1.0µM Pepstatin.
l Recommended final concentrations of phosphatase inhibitors:
l 1.0mM Na3VO4, 1.0 mM NaF.
