A precise method for the quantitation of proteins taking into account their amino acid composition
| Autor: | Hans-Joachim Horstmann |
|---|---|
| Rok vydání: | 1979 |
| Předmět: |
chemistry.chemical_classification
Chromatography Chemical Phenomena Total amino acids Hydrolysis Microchemistry Erythrocyte Membrane Biophysics Proteins Serum Albumin Bovine Cell Biology Fractionation Biochemistry Hydrolysate Amino acid Chemistry Ammonia chemistry.chemical_compound chemistry Amino acid composition Ninhydrin Humans Amino Acids Molecular Biology |
| Zdroj: | Analytical Biochemistry. 96:130-138 |
| ISSN: | 0003-2697 |
| DOI: | 10.1016/0003-2697(79)90564-5 |
| Popis: | A method for the quantitation of protein in biological material is described which gives the same response for all proteins irrespective of their amino acid composition. The method is based on the ninhydrin reaction of amino acids released after total acid hydrolysis of 5- to 20-μl solutions containing 1 to 100 μg of protein. The ammonia is released from the hydrolysate by diffusion and the amino acids are quantitated without fractionation using the continuous-flow system of an amino acid analyzer. Calibration is obtained with solutions of known amino acid content. The protein of a sample is calculated by multiplying the nanomoles of total amino acids found by a conversion factor F . F is the weight in micrograms of 1 nmol of the specific mixture of amino acid residues that the protein of the sample is composed of F has to be determined once for all further quantitations of the same material by quantitative amino acid analysis following standard procedures. By this method as little as 30 ng of protein per aliquot of hydrolysate analyzed can be determined. |
| Databáze: | OpenAIRE |
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