American Society of Hematology

Fig. 5.

$Fig. 5. Different-sized serum components are required for rosette formation. / (A) Sephacryl S-200 elution profile. Six-microliter fractions were collected, and the A280nm was determined for each. Fractions were pooled (S1-S3) and concentrated to the original volume of serum loaded. (B) SDS-PAGE profile of proteins in pooled fractions eluted from Sephacryl S-200. Aliquots of the whole serum (WS) and the pooled fractions (S1-S3) (up to 20 μg protein loaded, corresponding to the same relative volumes of each) were resolved by SDS-PAGE (10% acrylamide) as described in the legend to Figure 2. Molecular mass markers (M) are 19, 30, 46, 66, 97, and 220 kd from bottom to top, respectively. (C) Effect of the individual and recombined S-200 fractions on rosette formation. The rosetting assay was conducted as described in the legend to Figure 1, and the fractions were used at 15% (vol/vol) each for all experiments. The column was run using 2 different batches of serum, and the assay was conducted in duplicate for each batch (mean ± SD for 4 experiments).$

Different-sized serum components are required for rosette formation.

(A) Sephacryl S-200 elution profile. Six-microliter fractions were collected, and the A_280nm was determined for each. Fractions were pooled (S1-S3) and concentrated to the original volume of serum loaded. (B) SDS-PAGE profile of proteins in pooled fractions eluted from Sephacryl S-200. Aliquots of the whole serum (WS) and the pooled fractions (S1-S3) (up to 20 μg protein loaded, corresponding to the same relative volumes of each) were resolved by SDS-PAGE (10% acrylamide) as described in the legend to Figure 2. Molecular mass markers (M) are 19, 30, 46, 66, 97, and 220 kd from bottom to top, respectively. (C) Effect of the individual and recombined S-200 fractions on rosette formation. The rosetting assay was conducted as described in the legend to Figure 1, and the fractions were used at 15% (vol/vol) each for all experiments. The column was run using 2 different batches of serum, and the assay was conducted in duplicate for each batch (mean ± SD for 4 experiments).

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