Hello!
I want to prepare RIPA buffer to perform an ELISA essay.
Do you have a recipe to prepate RIPA buffer?
Thank you in advance.
Hello Benjamín Villasana-Salazar
You may prepare RIPA buffer (1X) as follows.
50mM Tris-HCL, pH 7.6
150mM NaCl
1% NP-40 or Triton X-100
0.5% Sodium deoxycholate
0.1% SDS
1mM EDTA
Add protease inhibitor cocktail fresh (1X) or use the following protease inhibitors such as Pepstatin A (1uM), Leupeptin (10uM) and PMSF (0.1mM) when you are about to perform cell lysis. Phosphatase inhibitors will be required to be added in the lysis buffer fresh only when phosphorylation states (activation states) are being investigated. The phosphatase inhibitors include Sodium fluoride (1mM) and Sodium orthovanadate (1mM). Perform cell lysis below 4 degree C using cold RIPA buffer.
Best.
Malcolm Nobre thank you!
Do you have a reference for this RIPA buffer recipe?
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