By Ronald L. Nagel
A set of with ease reproducible experimental and medical concepts for learning the molecular foundation of hemoglobin problems. those step by step tools make the most of a large choice of the newest analytical recommendations, together with x-ray crystallography, excessive functionality liquid chromatography, electrophoresis, and nuclear magnetic resonance. extra tools are provided for prenatal diagnostic research, the DNA prognosis of hemoglobin mutations, hemoglobin fluorescence, and the semisynthesis of hemoglobin.
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Additional resources for Hemoglobin Disorders: Molecular Methods and Protocols (Methods in Molecular Medicine)
2. The protein eluting at 1500 mL (~65% buffer B) corresponds to HbA. The fractions corresponding to this peak Purification and Molecular Analysis of Hb by HPLC 35 Fig. 1. 6 × 6 cm) at 4°C. 0. The column was equilibrated with buffer A, and a decreasing pH gradient of 0–100% buffer B over 20 CV was used for elution of the protein. Protein load: 25 mg. ). The following run takes about 25 to 26 h. 1. Pack the Q-Sepharose High Performance ion-exchange column at 4°C in a Pharmacia XK26/70 column according to the manufacturer’s directions.
And Gilliland, G. L. (1996) Crystallographic, molecular modeling, and biophysical characterization of the Valineβ67 (E11) → Threonine variant of hemoglobin. Biochemistry 35, 1935–1945. 42. Kavanaugh, J. , Chafin, D. , Rossi, G. , Kwiatkowski, L. , and Noble, R. W. (1995) Structure and oxygen affinity of crystalline desArg141 alpha human hemoglobin A in the T state. J. Mol. Biol. 248, 1136–1150. 43. Shih, D. T. , Luisi, B. , Perutz, M. , and Nagai, K. (1993). A mutagenic study of the allosteric linkage of His(HC3)146β in haemoglobin.
J. (1966) Abnormal human hemoglobins: separation and characterization of the a and b chains by chromatography, and the detereminatioin of two new variants, Hb Chesapeake and Hb J (Bangkok) J. Mol. Biol. 19, 91–108. 20. Brennan, S. O. (1985) The separation of globin chains by high pressure cation exchange chromatography. Hemoglobin 9, 53–63. 30 Wajcman Purification and Molecular Analysis of Hb by HPLC 31 3 Purification and Molecular Analysis of Hemoglobin by High-Performance Liquid Chromatography Belur N.