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. 2006 Aug 7;203(8):1985–1998. doi: 10.1084/jem.20060701

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Copyright © 2006, The Rockefeller University Press

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Figure 3. — Spleen and LN B cells from VH147 tg mice use similar endogenous L chains. (A) L chain cDNA was amplified and cloned from sorted VH147 tg spleen or LN anti-GPI B cells as described in Materials and methods. L chain assignment was determined by using the Ig BLAST and IMGT/V programs. The percentage of sequences using the given L chains is depicted from a total of n = 25 sequences each from the spleen and LN. (B) Relative affinity ELISA of VH147 tg B cells paired with endogenous L chains. Purified Ig from VH147 tg hybridomas was plated on serially diluted GPI-his-biotin bound to neutravidin-coated wells. Igs with a higher relative affinity for GPI will bind to lower concentrations of GPI than Igs with lower relative affinities. The graph represents the mean OD ± SEM from triplicate wells, normalized as the fraction of maximal binding. This experiment was repeated twice using at least 2 hybridomas for each L chain, yielding similar results.