The RB4 containing DTT (reducing agent) and 0.5 IPG buffer 3-10 non-linear (carrier ampholyte) exhibited more protein spots indicating better separation resolution. The precipitant containing TCA-acetone-DTT exhibited the highest protein recovery (82.2) demonstrating significant difference when compared with the other precipitants (p<0.05). Each of the saliva samples was subjected to the above experiments, carried out in triplicates. The efficiency of the different rehydration buffers was compared with respect to the resolution and focusing time taken to attain the maximum voltage. The precipitate can then be collected by centrifugation and the protein pellet is re-dissolved in a low salt buffer. The precipitate with the highest protein content was subsequently solubilized using different rehydration buffers (RB1, RB2, RB3 and RB4) before being subjected to the 2-DE. The efficiency of the precipitants was evaluated from protein content of the precipitate reflecting protein recovery. Five different protein precipitants: TCA, TCA-acetone-DTT, TCA-acetone-mercaptoethanol, acetone and alcohol were used to precipitate proteins of the saliva samples. Precipitants for salivary proteins and rehydration buffers for two-dimensional electrophoresis (2-DE) analysis were, respectively compared and evaluated. Protein_Precipitation_Method_for_Salivary_Proteins_and_Rehydration_Buffer_for_Two-Dimensional_Electrophoresis.pdf Popular answers (1) Johann P Klare Universitt Osnabrck Dear Md.Rezwanul Haque, quite often addition of a certain percentage of a stabilizing osmolyte like glycerol or sucrose (5-30, to be.
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