RIPA Lysis Buffer Calculator
Scale a source-defined RIPA lysis buffer at pH 8.0 from Tris, NaCl, EDTA, EGTA, nonionic detergent, deoxycholate, and SDS stocks.
Select a reviewed preset and enter the final volume to prepare.
| Reagent | Formula | Amount or action |
|---|---|---|
| 1 M Tris-HCl pH 8.0prepared aqueous stock | 1 mL | |
| 5 M sodium chlorideprepared aqueous stock | 3 mL | |
| 0.5 M EDTA solution, pH 8.0prepared aqueous stock | 200 µL | |
| 0.5 M EGTA pH 8.0prepared aqueous stock | 100 µL | |
| NP-40 or Triton X-100neat nonionic detergent | 1 mL | |
| 10% sodium deoxycholateprepared aqueous stock | 5 mL | |
| 10% SDSprepared aqueous stock | 1 mL | |
| Purified water | Bring the total solution volume to 100 mL. Do not add 100 mL of water; add water only until the total solution reaches 100 mL. |
How to prepare Standard RIPA pH 8.0
- Add the calculated Tris-HCl, NaCl, 0.5 M EDTA solution at pH 8.0, and EGTA stock volumes to less than the selected final volume of purified water.
- Add the calculated NP-40 or Triton X-100, sodium deoxycholate stock, and SDS stock volumes while mixing gently.
- Bring to the selected final volume with purified water and mix until uniform, avoiding excessive foaming.
- Add protease or phosphatase inhibitors fresh if required by your extraction protocol.
About This Buffer
RIPA refers to a family of detergent-containing lysis buffers used for extraction of cellular proteins, and exact formulations vary between protocols. This calculator scales one source-defined RIPA formulation at pH 8.0 containing Tris, NaCl, EDTA, EGTA, a nonionic detergent, sodium deoxycholate, and SDS. Protease or phosphatase inhibitors are not included as default scalable components.
Common questions
What is the final composition of this RIPA preset?
It contains 10 mM Tris-HCl pH 8.0, 150 mM NaCl, 1 mM EDTA, 0.5 mM EGTA, 1% nonionic detergent, 0.5% sodium deoxycholate, and 0.1% SDS.
Are protease and phosphatase inhibitors included?
No. They are optional fresh additions and must not be included automatically in the default recipe.
Can NP-40 and Triton X-100 be assumed to be chemically identical?
No. The preset permits the source-defined nonionic-detergent option, but the page should not claim that commercial mixtures are a single exact molecular species.
Sources and review notes
The source for the currently selected preset appears first. Recipe quantities remain tied to the cited article location.
Standard RIPA pH 8.0
Source location: RIPA buffer recipe — final concentrations are source-defined; displayed stock volumes are calculated from the registered stock strengths, with an independent composition cross-check. The source-defined EDTA stock is represented without inferring a salt or hydration state.
Preparation method: Source-defined final concentrations implemented with calculated volumes of defined stocks
- Detection of Retinoblastoma Protein Phosphorylation by Immunoblot Analysis PMC6613207 · open article
- Immunoblot Validation of Phospho-Specific Antibodies Using Lung Cancer Cell Lines PMC8034249 · open article