Biotinylated Human BCMA, Fc,Avitag (BC7-H82F0) is expressed from human 293 cells (HEK293). It contains AA Met 1 - Ala 54 (Accession # NP_001183.2).
Predicted N-terminus: Met 1
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 33.8 kDa. The protein migrates as 40-45 kDa under reducing (R) condition (SDS-PAGE) due to glycosylation.
Biotinylation of this product is performed using Avitag™ technology. Briefly, the single lysine residue in the Avitag is enzymatically labeled with biotin.
The biotin to protein ratio is 0.5-1 as determined by the HABA assay.
Less than 1.0 EU per μg by the LAL method.
>95% as determined by SDS-PAGE.
Lyophilized from 0.22 μm filtered solution in 50 mM Tris, 100 mM Glycine, 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:
- 4-8°C for 12 months in lyophilized state;
- -70°C for 3 months under sterile conditions after reconstitution.
Biotinylated Human BCMA, 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%.
Immobilized Human BAFF, Fc Tag (Cat. No. BAF-H4268) at 5 μg/mL (100 μL/well) can bind Biotinylated Human BCMA, Fc,Avitag (Cat. No. BC7-H82F0) with a linear range of 0.12-1.95 ng/mL (QC tested).
Immobilized Anti-Human BCMA Mab at 2 μg/mL (100 μL/well) can bind Biotinylated Human BCMA, Fc,Avitag (Cat. No. BC7-H82F0) with a linear range of 0.1-3 ng/mL (Routinely tested).
CAR阳性表达率检测（Evaluation of CAR expression ）
FACS Analysis of Anti-BCMA CAR Expression
293 cells were transfected with anti-BCMA-scFv and RFP tag. 2e5 of the cells were first stained with B. Biotinylated Human BCMA, Fc,Avitag (Cat. No. BC7-H82F0, 3 μg/ml) and C. biotinylated protein control, followed by FITC-conjugated streptavidin. A. Non-transfected 293 cells and C. biotinylated protein control were used as negative control. RFP was used to evaluate CAR (anti-BCMA-scFv) expression and FITC was used to evaluate the binding activity of Biotinylated Human BCMA, Fc,Avitag (Cat. No. BC7-H82F0).
FACS analysis of anti-BCMA CAR expression
Human T cells were transfected with anti-BCMA CAR and cultured for 3 days. Three days post-transfection, 1e6 cells were first incubated with 50ul Biotinylated Human BCMA, Fc,Avitag (Cat.No. BC7-H82F0, 8 μg/ml ), washed and then stained with PE Streptavidin and analyzed by flow cytometry. (Data are kindly provided by PREGENE Biopharma)
Authors: Berahovich R, et al.
Journal: Cancers (Basel) 2018
Application: Flow Cytometry
Tumor necrosis factor receptor superfamily member 17 (TNFRSF17) is also known as B-cell maturation protein (BCMA), CD antigen CD269, which is a member of the TNF-receptor superfamily. TNFRSF17 contains one TNFR-Cys repeat. TNFRSF17 is expressed in mature B-cells, but not in T-cells or monocytes. TNFRSF17 is receptor for TNFSF13B/BLyS/BAFF and TNFSF13/APRIL. TNFRSF17 promotes B-cell survival and plays a role in the regulation of humoral immunity. TNFRSF17 can activate NF-kappa-B and JNK.