Fig. 7.
Fig. 7. Immunoprecipitation analysis of allele-specific recombinant GPIIIa isoforms. / (A) Recombinant isoforms of GPIIb-IIIa complex were produced in CHO cells transfected either with Leu33, Leu33ΔLys611, Pro33, or Pro33ΔLys611 constructs of GPIIIa. Biotinylated cell lysates were immunoprecipitated with AB serum (lanes 1 and 5), anti–HPA-1a (lanes 2 and 6), anti–HPA-1b (lanes 3 and 7), and anti-Oea alloantibodies (lanes 4 and 8). Immunoprecipitates were analyzed on 7.5% SDS-PAGE under nonreduced conditions. (B) Electrophoretic mobilities of Pro33 and Pro33 ΔLys611 GPIIIa isoforms. Biotin surface-labeled GPIIb-IIIa complex derived from CHO transfectants was immunoprecipitated with mAb Gi5 and analyzed on 7.5% SDS-PAGE under nonreduced (NR) and reduced (RED) conditions. Lanes 1 and 5, Pro33; lanes 2 and 4, a mixture of Pro33 and Pro33 ΔLys611; and lanes 3 and 6, Pro33 ΔLys611 GPIIIa isoforms.

Immunoprecipitation analysis of allele-specific recombinant GPIIIa isoforms.

(A) Recombinant isoforms of GPIIb-IIIa complex were produced in CHO cells transfected either with Leu33, Leu33ΔLys611, Pro33, or Pro33ΔLys611 constructs of GPIIIa. Biotinylated cell lysates were immunoprecipitated with AB serum (lanes 1 and 5), anti–HPA-1a (lanes 2 and 6), anti–HPA-1b (lanes 3 and 7), and anti-Oea alloantibodies (lanes 4 and 8). Immunoprecipitates were analyzed on 7.5% SDS-PAGE under nonreduced conditions. (B) Electrophoretic mobilities of Pro33 and Pro33 ΔLys611 GPIIIa isoforms. Biotin surface-labeled GPIIb-IIIa complex derived from CHO transfectants was immunoprecipitated with mAb Gi5 and analyzed on 7.5% SDS-PAGE under nonreduced (NR) and reduced (RED) conditions. Lanes 1 and 5, Pro33; lanes 2 and 4, a mixture of Pro33 and Pro33 ΔLys611; and lanes 3 and 6, Pro33 ΔLys611 GPIIIa isoforms.

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