Ripa Lysis Buffer

SKU: orb348556

Description

Ripa Lysis Buffer

Images & Validation

• Application Notes:
  This product is ready-to-use as a working 1X solution and requires no further dilution. 1X 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 10E7 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 RIPA lysis buffer consists of 50 mM Tris HCl, 150 mM NaCl, 1.0% (v/v) NP-40, 0.5% (w/v) Sodium Deoxycholate, 1.0 mM EDTA, 0.1% (w/v) SDS and 0.01% (w/v) sodium azide at a pH of 7.4. This buffer was meticulously prepared using ultra pure 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: 1.0 mM Phenylmethylsulfonyl fluoride (PMSF) 10 μM Leupeptin 0.1 μM Aprotinin 1.0 μM Pepstatin Recommended final concentrations of phosphatase inhibitors: 1.0 mM Na3VO4 1.0 mM NaF

Key Properties

• Purity: 1X RIPA Lysis Buffer was aseptically filtered through a Millipore 0.22 micron filter into clean, pre-sterilized containers. The product was tested on trypticase soy agar for 24 hours, 48 hours and 72 hours and was found to be negative for bacteria.
• Conjugation: Unconjugated

Storage & Handling

• Storage: Store container at room temperature (18° to 26° C) prior to opening. Protect from light (store in the dark).
• Form/Appearance: Liquid (sterile filtered)
• Buffer/Preservatives: Preservative: 0.01% (w/v) Sodium Azide. Stabilizer: None; Buffer: See application note.
• Concentration: 1X
• Expiration Date: 12 months from date of receipt.
• Hazard Information: Non-Toxic
• Disclaimer: For research use only

Alternative Names

1X RIPA Lysis Buffer, 1X RIPA (Radio-Immunoprecipitation Assay) Lysis Buffer, RIPA Buffer

Comparison of myokines protein expression in the normal diet, normal diet + exercise, high fat diet, and high fat diet + exercise groups. (A) Myokines levels in skeletal muscle lysates were analyzed by western blot. (B–F) IL-7, IL-8, IL-6, CXCR2, and VEGF levels. Total proteins were extracted using RIPA lysis buffer (p/n orb348556) containing protease inhibitor cocktail, and 10 µg of protein was resolved by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and transferred to nitrocellulose membranes using the Trans-Blot Turbo Transfer System. Results represent the mean ± SE. *, p < 0.05 vs. the ND group by one-way ANOVA; †, p < 0.05 vs. the HFD group by one-way ANOVA. ND, normal diet; ND + Ex, normal diet + exercise; HFD, high fat diet; HFD + Ex, high fat diet + exercise.

Comparisons of protein expressions of (A) TrkB, (B) CaMkII, (C) AMPK, (D) PGC-1α, (E) FNDC5, and (F) β-amyloid in the cerebral cortex after the 12-week intervention. Total proteins were extracted using RIPA lysis buffer (p/n orb348556) containing a protease inhibitor cocktail, and 10 µg of protein was resolved by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and transferred to nitrocellulose membranes using the Trans-Blot Turbo Transfer System. (* p < 0.05 Significant difference as compared to Non-Ex group; # p < 0.05 Significant difference as compared to Saline group).