Which concentration of Rapigest should be used for protein extraction from a tissue sample?

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Juraj Lenco

RapiGest is quite expensive to be used for protein extraction. Is it really necessary to use this cleavable detergent? Why don't you try sodium deoxycholate instead? It will do the same job (protein extraction), it is compatible with BCA assay and trypsin digestion (trypsin loves it), it can be easy removed after digestion and it is inexpensive.

Murat Eravci

Hello Zahra, the folowing sentence is from the RapiGest SF manual (attached):

"The recommended concentration is 0.1% (w/v) RapiGest SF.

Hydrophobic proteins may require higher RapiGest SF concentrations (see note for recommendations for

working with proteolytic resistant or hydrophobic proteins)."

Best, Murat

Zahra Rashidbenam

Dear Dr. Murat, thank you for your comment. Yes I went through the guideline. However I think I will need to use 0.5% RapiGest as it was recommended by the supplier, to use 0.5% RapiGest 1:1 (Sample:Extraction buffer) if the sample does not need to be grounded into powder.

Dear Dr. Juraj,

Thank you for your suggestion. Yes, RapiGest is really expensive. I do like to replace it with something more reasonable in price. I am not sure if sodium deoxycholate is compatible with LCMS. As I know the only way to remove sodium deoxycholate is via dialysis, however it does not seem to be that efficient. Is there any other way to remove sodium deoxycholate ?

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Ali Sial

Hi Everyone,

I want to use Rapigest buffer to lyse the cells for for my proteomic sample preparation experiment. I have a new bottle of Rapigest but its expiry is September 2018. Can I still use it for my experiment? Will it work for lysis? or it will have any issue? Does anyone experience using buffer about to expire (such as Rapigest etc)?

Looking forward to hear soon. Thanks