A micromethod for the quantitation of cellular proteins in Percoll with the Coomassie brilliant blue dye-binding assay.
A simple procedure for the determination of cellular proteins in Percoll-containing samples is described. Percoll precipitated when particulate proteins were solubilized by dilution of the samples in a NaOH-Triton X-100 mixture. After centrifugation at high speed (12,000g), the supernatant was assayed for proteins with the Coomassie brilliant blue dye-binding assay. With an automatic spectrophotometer, 50-microliter aliquots gave a linear response between 0 and 3 micrograms of bovine serum albumin. After a fivefold dilution in the alkali-detergent mixture, proteins in samples containing up to at least 60% Percoll can be accurately quantitated on a standard curve prepared in the absence of Percoll. Because the sensitivity of the assay was better than 100 ng, the procedure outlined in this paper can also be used as a general protein micromethod.
D000818:Animals; D001322:Autoanalysis; D006801:Humans; D008264:Macrophages / Q000032:analysis*; D011205:Povidone; D011506:Proteins / Q000032:analysis*; D051381:Rats; D012394:Rosaniline Dyes; D012710:Serum Albumin, Bovine / Q000032:analysis; D012822:Silicon Dioxide; D013053:Spectrophotometry; D013223:Statistics as Topic
D011506:Proteins; D012394:Rosaniline Dyes; D012710:Serum Albumin, Bovine; C016039:Percoll; D012822:Silicon Dioxide; D011205:Povidone; C004692:coomassie Brilliant Blue