Hi, I wanted to isolate proteins using trizol method. During performing the method, i was unable to dissolve the pellet in buffer comprising 0.3 M Gn-HCl, ethanol, 2.5% glycerol. However i vortexed alot and was able to dissolve tiny pellet. Despite i see more undissolved pellet, proceeded with following steps. At the end the protein pellet is solublised completely in solubilization buffer, 100 mM Tris-Hcl, pH 8.0 contains 2% SDS, kept it at 55 degree, 5 minutes.
Can i use this protein solution for down stream applications, despite the protein pellet was undissolved completely in buffer, 0.3 M Gn-HCl, ethanol, 2.5% glycerol?
TIA
A protein concentration of the last solution must be determined to verify if your protein is solubilized on it. All procedures must be based on this last protein concentration. Another fact must be considered. After all procedures you submitted it, the protein properties are intact yet?
Roseli A Gomes Thank you for your response. Solubilized protein concentration determined and it is 1 mg/ml. I am not sure about protein properties are intact or not?
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