Fig. 2.
Fig. 2. Sequential immunoprecipitation experiments of metabolically labeled lymphoblastoid cells with antibodies specific for human ferritin H-chain (αH) and L-chain (αL). (A) Control cells (C1) and Verona-1 cells (Ver-1) were metabolically labeled with35S-methionine, lysed, and 2 × 106 cpm of the soluble fraction of homogenates precipitated with saturating amounts of anti-H chain antibody. The soluble fraction was then precipitated again with saturating amounts of anti–L-chain antibody. The precipitates were analyzed on SDS-PAGE under denaturing conditions and exposed to autoradiography. (B) Verona-1 cells were metabolically labeled with35S-methionine and immunoprecipitated as in (A), the pellets were resuspended in 4 mol/L urea, and separated on 6% polyacrylamide gels containing 4 mol/L urea under conditions that disrupt antibody antigen interactions without affecting ferritin structure. (C) As in (B), except that cells were metabolically labeled with 59FeNTA.

Sequential immunoprecipitation experiments of metabolically labeled lymphoblastoid cells with antibodies specific for human ferritin H-chain (αH) and L-chain (αL). (A) Control cells (C1) and Verona-1 cells (Ver-1) were metabolically labeled with35S-methionine, lysed, and 2 × 106 cpm of the soluble fraction of homogenates precipitated with saturating amounts of anti-H chain antibody. The soluble fraction was then precipitated again with saturating amounts of anti–L-chain antibody. The precipitates were analyzed on SDS-PAGE under denaturing conditions and exposed to autoradiography. (B) Verona-1 cells were metabolically labeled with35S-methionine and immunoprecipitated as in (A), the pellets were resuspended in 4 mol/L urea, and separated on 6% polyacrylamide gels containing 4 mol/L urea under conditions that disrupt antibody antigen interactions without affecting ferritin structure. (C) As in (B), except that cells were metabolically labeled with 59FeNTA.

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