To produce this hybridoma mice transgenic HLA-B27 and human β-2 microglobulin were immunized with a preparation of HFE made in recombinant bacteria, refolded with human β-2 microglobulin. Characterisation beyond data in B.J. Haematol 103, 931, 1998 shows that JB1 reacts predominantly with aggregated denatured HFE produced in bacteria, but does not bind folded recombinant HFE produced in CHO cells. In comparison with polyclonal sera raised to the C terminus or the α 3 region of HFE JB1 stains frozen sections of the tissues well, but reacts very weakly with cells transfected with the HFE gene in vitro (usually only visible by confocal microscopy). JB1 does not stain paraffin fixed sections and will not work in western blots.
Immunogen
Bacterial recombinant HFE protein
Culture Medium
RPMI 1640, 10% FBS, 1% HAT
Subculture Routine
Maintain cells at 3-5x100,000 cells/ml which is usually a split ratio of 1:2. Saturation density os 5 x 100,000 to 1 x 1000,000.
Other Notes
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