Copurification of P6, MRP8, and MRP14 from human granulocytes and separation of individual proteins.
A method is described for purification of P6, MRP8, and MRP14, three calcium-binding proteins assigned to the S100 protein family. The purification procedure included preparation of human granulocytes, ammonium sulfate precipitation, and anion-exchange chromatography and resulted in the copurification of P6, MRP8, and MRP14. Individual proteins were separated by either preparative isoelectric focusing or preparative SDS-PAGE. The procedure was carried out in the course of 4 days and yielded several milligrams of essentially pure P6, MRP8, and MRP14 in either native or denatured form.
van den Bos
C
C
Institute of Experimental Dermatology, von Esmarchstrasse 56, Muenster, 48159, Germany. cvandenbos@osiristx.com
Rammes
A
A
Vogl
T
T
Boynton
R
R
Zaia
J
J
Sorg
C
C
Roth
J
J
D000595:Amino Acid Sequence; D000943:Antigens, Differentiation / Q000097:blood / Q000737:chemistry / Q000302:isolation & purification*; D002135:Calcium-Binding Proteins / Q000097:blood / Q000737:chemistry / Q000302:isolation & purification*; D040501:Calgranulin A; D040502:Calgranulin B; D002478:Cells, Cultured; D002852:Chromatography, Ion Exchange; D004591:Electrophoresis, Polyacrylamide Gel; D006098:Granulocytes / Q000378:metabolism*; D006801:Humans; D007525:Isoelectric Focusing; D008969:Molecular Sequence Data; D009418:S100 Proteins
D000943:Antigens, Differentiation; D002135:Calcium-Binding Proteins; D040501:Calgranulin A; D040502:Calgranulin B; D009418:S100 Proteins; C114242:calcium-binding protein p6, human