Biochimica Et Biophysica Acta 625 Bba 26182 Use of T H E Zonal Centrifuge to Separate Particles Containing Transplantation Antigen

نویسندگان

  • RAYMOND A. POPP
  • DIANA M. POPP
  • NORMAN G. ANDERSON
  • L. H. ELROD
چکیده

Subcellular particles of homogenates of mouse spleen were separated at various centrifugal forces between 5oo0 • lO 6 and 30 ooo. lO 6 o~t in the zonal centrifuge. Each fraction was analyzed for transplantation antigen, ATPase, nonspecific esterase and alkaline phosphatase. Under appropriate conditions, zonal centrifugation proved tobe an efficient procedure to separate particles containing high concentrations of transplantation antigen. After a centrifugal force of 5000' lO 6 w2t, the antigenic particles were located in the lO-12 % sucrose region, but they sedimented into denser sucrose particles and became more disperse when subjected to larger total centrifugal forces. The density of the particles in CsC1 was about 1.2 g/cm 3. Transplantation antigen was solubilized by autolysis; the antigen was eluted with the exclusion volume during gel filtration on Sephadex G-2oo.

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تاریخ انتشار 2002