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Your Position: Home > MABSol® Biotin Labeled Proteins > CD19 > Biotinylated Human CD19 Protein With C-Fc Tag

Biotinylated Human CD19, Fc Tag, ultra sensitivity (primary amine labeling)

  • Synonym
    CD19,B4,CVID3,MGC12802
  • Source
    MABSol® Biotinylated Human CD19, Fc Tag (CD9-H8259) is expressed from human HEK293 cells. It contains AA Pro 20 - Lys 291 (Accession # AAH06338). It is the biotinylated form of Human CD19 Protein, Fc Tag (Cat # CD9-H5259).
    Predicted N-terminus: Pro 20
  • Molecular Characterization
    CD19(Pro 20 - Lys 291) AAH06338

    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.

  • Biotinylation
    The primary amines in the side chains of lysine residues and the N-terminus of the protein are conjugated with biotins using standard chemical labeling method. A standard biotin reagent (13.5 angstroms) is used in this product.
  • Biotin:Protein Ratio
    The biotin to protein ratio is 2.5-3.5 as determined by the HABA assay.
  • Endotoxin
    Less than 1.0 EU per μg by the LAL method.
  • Purity

    >95% as determined by reduced SDS-PAGE.

  • Formulation

    Lyophilized from 0.22 μm filtered solution in PBS, pH7.4. Normally trehalose is added as protectant before lyophilization.

    Contact us for customized product form or formulation.

  • Reconstitution

    Please see Certificate of Analysis for specific instructions.

    For best performance, we strongly recommend you to follow the reconstitution protocol provided in the CoA.

  • Storage

    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 is observed after storage at:

    1. 4-8°C for 12 months in lyophilized state;

    2. -70°C for 3 months under sterile conditions after reconstitution.

SDS-PAGE
Biotinylated Human CD19, Fc tag (Cat. No. CD9-H8259) SDS-PAGE gel

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%.

Bioactivity
Biotinylated Human CD19, Fc tagBiotinylated Human CD19, Fc tag (Cat. No. CD9-H8259) ELISA bioactivity

Immobilized FMC63 MAb at 2 μg/mL (100 μL/well) can bind Biotinylated Human CD19, Fc Tag (Cat. No. CD9-H8259) with a linear range of 0.078-1.25 μg/mL (QC tested).

Evaluation of CAR expression
FACS Analysis of Anti-CD19 CAR Expression
Evaluation of CAR expression

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, 10ug/ml ) . C. Recombinant biotinylated human CD19 (Cat. No. CD9-H8259, 10ug/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) .

FACS Analysis of Anti-CD19 CAR Expression
Evaluation of CAR expression

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.)

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