How much to dilute whole cell liver tissue extract for micro Lowry protein concentration determination?
You don't provide enough information. The answer would depend on the volume used to homogenize the liver, as well as the dynamic range of the particular version of the Lowry assay. I suggest that you prepare a set of serial dilutions and assay them all to make sure of getting at least one in the right range.
I agree with the above. We would need more information.
As a reference:
I've found that lysis of mouse liver with Laemmli's buffer (2% SDS, 33mM Tris HCl @ pH 6.8, 5% Glycerol) liberated ~30% of tissue mass as detectable protein by BCA assay (akin to Lowry, I think) using a BSA standard curve, when lysed via sonication at Tissue:Buffer ratios of 1mg:10-30ul. Under similar lysis condidtions RIPA has a ~20% yield. Omitting the sonication step can reduce extraction efficiency by both buffers by up to 50%.
You might be able to use this data to get an estimate of the needed dilution, but it would be better to determine it empirically, as suggested above.
Thank you for replays. I used 1:4 ratio tissue: buffer. I used than macro lowry for protein determination and i went rather well. I got concentrations around 10 ug/ul. I used hepes buffer pH7,4 as homogenization buffer with triton x 100, glycerol and cocktail of protease and phosphatase inhibitors.
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