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Application notes
Himac Application No. 57 – Separation of Lipoprotein from Human Serum
Himac CP Series Ultracentrifuge and P100AT2 Fixed Angle Rotor
Human serum lipoprotein is thought to have something to do with atherosclerosis or hyperlipidemia and recently the association is investigated in various ways. We tried to separate lipoprotein rapidly from human serum by high performance preparative ultracentrifuge CP100 with maximum rotating speed of 100, 000rpm (maximum centrifugal force of 803, 000 X g).
1. Models used
Centrifuge : Hitachi high performance preparative
ultracentrifuge CP100
Rotor: P100AT2
(Fixed-angle rotor)
Tube: 4.7PC
thick-walled tube (Single use only)
Cap: B - Ti lid (Be sure to use)
2. Methods and results
Fat Red 7B is added for recognition of the results easily.1)
Soln. A (1.006g/cm3) : Dissolve 11.4g of NaCl and 0.1g of EDTA-Na2 with 500ml of distilled water (DW) and add 1ml of 1N NaOH. Then add DW up to 1000ml and add 3ml of DW. (NaCl : 0.195mol)
Soln. B (1.182g/cm3) : Dissolve 24.98g of NaBr to 100ml of soln.A. (NaCl : 0.195mol, NaBr : 2.44mol) Soln. C (1.478g/cm3) : Dissolve 78.32g of NaBr to 100ml of soln.A. (NaCl : 0.195mol, NaBr : 7.65mol)
Reference
1)Bronzert T. J. and Brewer H. B. Clin. Chem., 23 , 2089 (1977).
Product information Himac Ultracentrifuge