This protein carries a human IgG1 Fc tag at the C-terminus, followed by a Avi tag (Avitag™).
The protein has a calculated MW of 103.9 kDa. The protein migrates as 115-120 kDa under reducing (R) condition (SDS-PAGE) due to glycosylation.
>95% as determined by SDS-PAGE.
Lyophilized from 0.22 μm filtered solution in Tris with Glycine, Arginine and NaCl, pH7.5. 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:
Biotinylated Human Siglec-2, Fc,Avitag 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 anti-CD22-scFv and RFP tag. 2e5 of the cells were first stained with B. Biotinylated Human Siglec-2, Fc,Avitag (Cat. No. SI2-H82F8, 10 μg/mL) and C. Biotinylated Protein Control, followed by FITC Streptavidin. A. Non-transfected 293 cells and C. Biotinylated Protein Control were used as negative control. RFP was used to evaluate CAR (anti-CD22-scFv) expression and FITC was used to evaluate the binding activity of Biotinylated Human Siglec-2, Fc,Avitag (Cat. No. SI2-H82F8).
Human T cells were lentivirally transduced with anti-CD19 CAR and cultured for 3 days. Three days post-transduction, 1e6 cells were first incubated with 100 μL Biotinylated Human Siglec-2, Fc,Avitag (Cat. No. SI2-H82F8, 10 μg/mL), washed and then stained with PE Streptavidin. Non-transduced T cells were used as a control for gating of CAR expression. (Data are kindly provided by Cellyan Therapeutics Co. Ltd.)
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