I am looking for the protocol of RIPA buffer in the internet and found many different kinds of RIPA buffer, some have NaF and some without, does anyone know the function of NaF in the RIPA buffer?
Thank you!
Hi Zhengrong
Look at link and article
http://www.protocol-online.org/biology-forums/posts/8747.html
Sodium fluoride is a phosphatase inhibitor. We use it in most cell lysis buffers and it does not present a problem.
Sodium fluoride, as well as Sodium PyroPhosphate and Sodium Ortovanadate, is usually added in protein lysis buffers (such as RIPA, JS or E1A) to inactivate endogenous phosphatases and protect protein phosphorylations. These inhibitors are therefore necessary whenever you are interested in the WB analysis of phospho-proteins.
اHello
yes , always used NaF and Na3Vo4 to preserve the protein phosphorylation state in cells, cell lysates, and protein kinase (PK) assays. These inhibitors should ideally be present in all buffers used in the purification of PKs .
Good Luck
Hello,
Does anyone know if NaF and Na3Vo4 is "selective" for phosphatase?
Does it inhibit phospholipase a2 activity?
I didn't find any information on.
Thank you!
Hello,
NaF is a ser/thr phosphatase
Na3VO4 is a Tyr phosphatase ans alkaline phosphatase
Inhibitors.
As listed in thermoFisher webpage.
https://www.thermofisher.com/us/en/home/life-science/protein-biology/protein-biology-learning-center/protein-biology-resource-library/pierce-protein-methods/protease-phosphatase-inhibitors.html#protease