Hypoosmotic dialysis and ultrafiltration technique for preparation of mammalian hemoglobin: a comparison of three species.
Dialysis of human, bovine, and ovine red blood cells using a hypotonic solution and a commercial kidney dialysis unit, followed by ultrafiltration through 0.1 micron pore hollow-fibers provides an easily managed method for isolation of lipid-free hemoglobin (LFHB). High performance liquid chromatography (HPLC) analysis of hemoglobin indicated 99% protein purity. SDS polyacrylamide gel electrophoresis analysis demonstrated that LFHB migrates as a single band. Processing one-half liter of packed red blood cells requires approximately 10 hours and resulting in an average of 93% hemoglobin recovery.
Sheffield
C L
CL
Veterinary Toxicology and Entomology Research Laboratory, United States Department of Agriculture, College Station, Texas 77841.
Spates
G E
GE
DeLoach
J R
JR
D000818:Animals; D002417:Cattle; D002851:Chromatography, High Pressure Liquid; D003956:Dialysis; D004591:Electrophoresis, Polyacrylamide Gel; D004912:Erythrocytes / Q000648:ultrastructure; D006454:Hemoglobins / Q000302:isolation & purification*; D006801:Humans; D008055:Lipids / Q000097:blood; D008855:Microscopy, Electron, Scanning; D009997:Osmotic Pressure; D012756:Sheep; D013045:Species Specificity; D014462:Ultrafiltration
D006454:Hemoglobins; D008055:Lipids