membrane protein purification
Posted 16 February 2010 - 01:03 PM
I'm trying to purify one membrane protein, which is solubilized in RIPA buffer containing 1% triton x-100, Na DOC, Nacl, EDTA, SDS and Mgcl2.
The problem is when I try to quantify the samples by bradford, the cuvettes are with intense blue!!! even the cuvettes of BSA for the standard courb.
I'm wondering which compound of RIPA that interfere with bredford!! I tried also by turbidometric method TCA; all the samples are precipitated before the quantification by spectrofotometer.
Please, if you have any suggestions,
waiting for reply soon
Posted 16 February 2010 - 03:35 PM
Posted 16 February 2010 - 03:41 PM
I will try today to change this parameter.
Posted 16 February 2010 - 03:54 PM
In fact, I read in the literature some protocols using RIPA with the same components as mine, but, i don't know how they quantified proteins!!
thanks for advice
Posted 17 February 2010 - 08:02 AM
genius does what it must
i do what i get paid to do
Posted 17 February 2010 - 03:34 PM
I'm thinking to use the same ripa buffer diluted since I read it also in the "Thermo scientific total protein assays", dilution 1:10. without changing triton 1% or deoxycholate, I will see if it works, to quantify my samples!!!
thanks for interaction