| First Author | Wixler V | Year | 1994 |
| Journal | J Immunol Methods | Volume | 171 |
| Issue | 1 | Pages | 121-30 |
| PubMed ID | 8176233 | Mgi Jnum | J:18092 |
| Mgi Id | MGI:66111 | Doi | 10.1016/0022-1759(94)90235-6 |
| Citation | Wixler V, et al. (1994) Isolation and quantification of class 1 MHC gene mutants in mouse T cells by immunoselection with a magnetic cell sorter (MACS). J Immunol Methods 171(1):121-30 |
| abstractText | A method is described for determining the frequency of cells with a mutation in an autosomal gene coding for a membrane protein. Using a monoclonal antibody to H-2Kk surface antigen and magnetic cell separation (MACS) more than 10(4)-fold enrichment of the H-2Kk negative population was achieved, as tested with artificial mixtures containing a known number of antigen-negative cells. After a second magnetic sorting mutant frequencies as low as 10(-6) could be measured. The number of clonogenic mutants was evaluated by limiting dilution cloning and verification of the mutant phenotype by FACScan (flow cytometry) analysis in a representative number of clones. The spontaneous frequency of H-2Kk deficient mutants was 0.80 x 10(-6), and this increased after irradiation with 6 Gy X rays to 3.38 x 10(-6) within the next 8 weeks. About 50 mutant clones were screened for the presence of other class 1 antigens on the cell surface by FACScan analysis. All mutants continued to express other class 1 antigens. |
