Purification of histone H1 polypeptides by high-performance cation-exchange chromatography
| Autor: | J.D. Paulis, M.J. Fritzler, J. Gohill |
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| Rok vydání: | 1990 |
| Předmět: |
Ion chromatography
Size-exclusion chromatography Enzyme-Linked Immunosorbent Assay Fractionation Thymus Gland Procainamide Sodium Chloride Biochemistry High-performance liquid chromatography Analytical Chemistry Matrix (chemical analysis) Histones Histone H1 medicine Animals Chymotrypsin Lupus Erythematosus Systemic Bromosuccinimide chemistry.chemical_classification Chromatography medicine.diagnostic_test Organic Chemistry Thrombin General Medicine Enzyme chemistry Immunoassay Cattle Chromatography Liquid |
| Zdroj: | Journal of chromatography. 502(1) |
| Popis: | Calf thymus histone 1 (H1) was cleaved by chemical and enzymatic methods and the resulting polypeptides were fractionated by high-performance cation-exchange. Up to 1 mg of H1 polypeptides were loaded onto a 50 × 5 mm I.D. cation-exchange column and fractionated to greater than 95% purity in less than 30 min. This is the first report on the separation of H1 polypeptides by a strong cation-exchange matrix. In addition, the high-performance cation-exchange chromatography protocol represents a significant decrease in fractionation time when compared to conventional ion-exchange and gel filtration chromatography. The utility of this procedure is shown when the H1 peptides purified by the protocol were used to define antigenic domains of H1 band by procainamide-induced lupus and idiopathic systemic lupus erythematosus. The majority of the sera tested by enzyme-linked immunoassay (ELISA) reacted to the C-terminal peptides of H1 indicating this to be the major antigenic domain of H1. |
| Databáze: | OpenAIRE |
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