By Hernandes M. E.
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Standard protein solution, 3 mg/ml Weigh out dry protein and prepare a stock solution at a concentration of 3 mg/ml in the same solvent as used for the sample protein. Store up to 3 months at -20°C. To prepare calibration standard solutions, dilute the stock solution in solvent to give the desired final concentrations for the standard curve. Bovine serum albumin (BSA, fraction V; Sigma) is frequently used for a protein standard solution. 61 for a 1 % ( w h ) solution. For quantitution of a purijied or partially purified protein, ifpossible, the protein standard should have an aromatic amino acid content similar to that of the sample protein.
However, with a protein standard whose aromatic amino acid content is similar to that of the sample, intrinsic fluorescence can be used for quantitation (Hawkins and Honigs, 1987). , adjacent protonated acidic groups in a protein molecule will quench tryptophan fluorescence (Freifelder, 1982). Critical Parameters and Troubleshooting A I-cm path length quartz cuvette is most often used to make absorbance measurements. , from Hellma Cells or Beckman); these shorter path length cuvettes allow higher concentrations of protein solutions to be measured.
7. Transfer reaction vessel to a 55°C water bath and incubate for exactly 19 min. 8. Transfer reaction back to the 37°C bath, insert pH electrode, and read pH at exactly 20 min. The p H at 20 min is X . 9. 56 (X) 10. Repeat steps 5 to 9 for test proteins and calculate 9% digestibility as per equation in step 9. REAGENTS AND SOLUTIONS Use deionized or distilled water in all recipes and protocol steps. For common stock solutions, see APPENDZX 2A; ,for suppliers, see SUPPLIERS APPENDIX. 0 with NaOH.
Ae -codimension of germs of analytic curves by Hernandes M. E.