| Description |
A genetically defined, serologically identified antigen of the rat lymphocyte membrane (AgF-1) has been isolated. Viable spleen and lymph node cells, prepared by Ficoll-Hypaque density centrifugation from Fisher rats, were radioiodinated using soluble lactoperoxidase. Extracts, obtained with Nonidet P-40 were shown to contain numerous radio-labeled proteins, including cell-surface globulin. AgF-1 was isolated from these extracts by precipitation with a highly specific alloantibody in conjunction with xenospecific anti-globulin antibody and polyethylene glycol (PEG). The use of PEG greatly increased the efficiency of the double antibody technique. The putative antigenic peak was eluted from sodium dodecyl sulfate polyacrylamide gels (SDS-PAGE) and specific antigenic activity was recovered. Removal of the SDS from there eluates was achieved by equilibration with urea and passage over an anion exchange resin. Renaturation, as evidenced by specific inhibition of complement-mediated cytotoxicity, occurred upon the removal of urea by dialysis. The molecular weight of the purified antigen was estimated to be 35-40,000 daltons by SDS-PAGE and was unaffected by reduction with 2-mercaptoethanol. Amino acid composition was roughly similar to those reported for the major histocompatibility antigens of the rat and other species. |
| OCR Text |
Show UNIVERSITY OF UTAH GRADUATE SCHOOL SUPER VISOR Y COMMITTEE APPROV AL of a dissertation submitted by Phillip B. C. Williams I have read this dissertation and have found it to be of satisfactory quality for a doctoral d ree ' r j Date *1/ I ( q I �;,�j , � 7[;> eWitt __ Chairman, Supervisory Committee I have doctora �"'I"". Date have I have read this dissertation and doctoral degree. "FI4 -" _ J Dare found it to be of satisfactory quality for a � ��� �� C�-t""'� � � 4� �� - __ ___ Swanson Member. Supervisory Committee I have read this dissertation and have found it to be of satisfactory quality for , doctoral degree. " -_ '1 Date ( '�-') /,.',: '/(., Ha�i a �fii�� L.k:;::U::i"� Member, Supervi�ory Committee /--'. I have read this dissertation d;jy �! 7f Dare and ,--:' /i l / �{f � � satisfactory quality for a have found / _ S_/( tJ.1i{ � ��g"'-I_'I /" -''''-)6 Lawrence B."'- Sandberg Member, Supervisory Committee __ UNIVERSITY OF UTAH GRADUATE SCHOOL FINAL READING To t he Graduate Council of the University of I have read the dissertation of final fonn and have found " , ___ _ _ that (1) APPROVAL Utah: Pbillip,_�,. __ C, Jii.lli,am,s in its and bibliographic style are , ____ ___ its format, citations, consistent and acceptahle; (2) its illustrative materials including figures, tables, and charts are in place; and (3) th e final manuscript is satisfactory to the Supervisory Committee and is rendy for submissjon to I / ,', I, �'I� f ;!= -- / �// - -'" 'n; __ _ the Graduate School. f.:� (J..�_�'L<c.Lj�_� Charles W. __ _ DeWitt Member, Supervisory Committet! Approved for the Maj or -------' -- ·1ll.-�i.' Erns <- l.(, J. Department �! ----- - --- - ------- Eichwald Chairman /Dean Approved for thr (;raduat(�.. Coutl(�il Sterling M. Dean McMurrin of the Graduate School |
