Crude spectrin extraction from reticulocyte-rich blood samples.
Crude spectrin was extracted from the isolated red cell ghosts with low ionic strength buffer at 37 degrees C for 30 min. No significant alterations in crude spectrin extractability in wide range of patients with various hematologic diseases were observed. However, blood samples characterized by elevated reticulocytosis provided crude extracts with increased amount of non-heme membrane skeletal proteins. The presence of ribose in the crude spectrin extracts obtained from reticulocyte-rich blood samples indicates also the presence of nucleic acids which causes the shift of protein peak in the extract from 280 nm towards lower wavelengths. A model experiment with a normal crude spectrin extract mixed with various amounts of RNA allowed us to obtain the correction curve which served for determination of non-heme protein (crude spectrin) extractability.
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Marík
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Kodícek
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Brabec
D001797:Blood Protein Electrophoresis; D004591:Electrophoresis, Polyacrylamide Gel; D004910:Erythrocyte Membrane / Q000032:analysis*; D006402:Hematologic Diseases / Q000097:blood; D006801:Humans; D007202:Indicators and Reagents; D012313:RNA / Q000097:blood; D012156:Reticulocytes / Q000032:analysis*; D013049:Spectrin / Q000302:isolation & purification*; D013056:Spectrophotometry, Ultraviolet
D007202:Indicators and Reagents; D013049:Spectrin; D012313:RNA