Fig. 1.
Fig. 1. Purification of recombinant granzymes and NK cell-derived perforin. / (A) Recombinant granzymes were produced and purified (0.5 μg), and equivalent amounts (1 μg) of rGzmA, rGzmB, rGzmBSer183Ala, rGzmC, and rGzmCSer184Ala were separated on an SDS-PAGE reducing gel and visualized by silver staining. (B) Western blots. NK cell line lysates (25 μg) and 0.5 μg of partially purified perforin, rGzmA, rGzmB, rGzmBSer183Ala, rGzmC, and rGzmCSer184Ala were electrophoresed on SDS-PAGE gels. Western blot analysis was performed using polyclonal rabbit anti-mouse GzmA, B, and C and the rat antihuman perforin antibody (1:2000 each).

Purification of recombinant granzymes and NK cell-derived perforin.

(A) Recombinant granzymes were produced and purified (0.5 μg), and equivalent amounts (1 μg) of rGzmA, rGzmB, rGzmBSer183Ala, rGzmC, and rGzmCSer184Ala were separated on an SDS-PAGE reducing gel and visualized by silver staining. (B) Western blots. NK cell line lysates (25 μg) and 0.5 μg of partially purified perforin, rGzmA, rGzmB, rGzmBSer183Ala, rGzmC, and rGzmCSer184Ala were electrophoresed on SDS-PAGE gels. Western blot analysis was performed using polyclonal rabbit anti-mouse GzmA, B, and C and the rat antihuman perforin antibody (1:2000 each).

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