This protein carries a human IgG1 Fc fragment at the C-terminus. The protein has a calculated MW of 56.3 kDa. The protein migrates as 56-66 kDa on a SDS-PAGE gel under reducing (R) condition due to glycosylation.
>95% as determined by reduced SDS-PAGE.
Lyophilized from 0.22 μm filtered solution in PBS, pH7.4. Normally trehalose is added as protectant before lyophilization.
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Please see Certificate of Analysis for specific instructions.
For best performance, we strongly recommend you to follow the reconstitution protocol provided in the CoA.
For long term storage, the product should be stored at lyophilized state at -20°C or lower.
Please avoid repeated freeze-thaw cycles.
No activity loss was observed after storage at:
4-8°C for 12 months in lyophilized state;
-70°C for 3 months under sterile conditions after reconstitution.
Biotinylated Human CD19, Fc Tag on SDS-PAGE under reducing (R) condition. The gel was stained overnight with Coomassie Blue. The purity of the protein is greater than 95%.
293 cells were transfected with FCM63-scFv and RFP tag . 2x105 of the cells were first incubated with A. Biotinylated protein control. B. Recombinant biotinylated human CD19 (Cat. No. CD9-H8259, 10 μg/ml ) . C. Recombinant biotinylated human CD19 (Cat. No. CD9-H8259, 10 μg/ml) and FMC63(Mouse anti-CD19 antibody). FITC Streptavidin was used to analyse with FACS. RFP was used to evaluate CAR(FMC63-scFv) expression and FITC was used to evaluate the binding activity of recombinant biotinylated human CD19 (Cat. No. CD9-H8259) .
Human T cells were lentivirally transduced with anti-CD19 CAR and cultured for 11 days. Eleven days post-transduction, 1e6 cells were stained for the expression of CD3 and anti-CD19 CAR with FITC anti-human CD3 antibody and biotinylated human CD19 (Cat. No. CD9-H8259) followed by PE-conjugated streptavidin, respectively. A. Non-transduced T cells were used as a control for gating of CAR expression. (Data are kindly provided by Beijing Bowei Huaen Medical Technology Co. Ltd.)
Authors: MacLeod DT, et al.
Journal: Mol Ther 2017
Application: Flow Cytometry
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