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Your Position: 홈 > Protein > CD3E & CD3D > CDD-H82W0

Biotinylated Human CD3E&CD3D Heterodimer Protein, Fc,His Tag&Fc,Flag Tag, ultra sensitivity (primary amine labeling) (MALS verified)

INTRODUCTION
QC & Assay Protocol
Citation & Background
  • Synonym
    CD3E & CD3D,CD3 delta & CD3 epsilon
  • Source
    MABSol® Biotinylated Human CD3E&CD3D Heterodimer Protein, Fc,His Tag&Fc,Flag Tag, primary amine labeling (CDD-H82W0) is expressed from human HEK293 cells. It contains AA Asp 23 - Asp 126 (CD3E) & Phe 22 - Ala 105 (CD3D) (Accession # NP_000724.1 (CD3E) & NP_000723.1 (CD3D)).
    Predicted N-terminus: Asp 23 (CD3E) & Phe 22 (CD3D)
    Request for sequence
  • Molecular Characterization
    CD3E & CD3D Structure

    The subunit CD3E carries a human IgG1 Fc fragment at the C-terminus, followed by a polyhistidine tag. The subunit CD3D carries a human IgG1 Fc fragment at the C-terminus, followed by a flag tag. The protein has a calculated MW of 39.2 kDa (CD3E) and 37.5 kDa (CD3D) . The protein migrates as 45-55 kDa on a SDS-PAGE gel when calibrated against Star Ribbon Pre-stained Protein Marker under reducing (R) condition due to glycosylation.

  • Labeling
    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.
  • Protein Ratio
    Passed as determined by the HABA assay / binding ELISA.
  • Endotoxin
    Less than 1.0 EU per μg by the LAL method.
  • Purity

    >95% as determined by SDS-PAGE.

    >90% as determined by SEC-MALS.

  • Formulation

    Lyophilized from 0.22 μm filtered solution in PBS, pH7.4 with trehalose as protectant.

    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.

    This product is stable after storage at:

    1. -20°C to -70°C for 12 months in lyophilized state;
    2. -70°C for 3 months under sterile conditions after reconstitution.
SDS-PAGE
CD3E & CD3D SDS-PAGE

Biotinylated Human CD3E&CD3D Heterodimer Protein, Fc,His Tag&Fc,Flag Tag, primary amine labeling on SDS-PAGE under reducing (R) condition. The gel was stained with Coomassie Blue. The purity of the protein is greater than 95% (With Star Ribbon Pre-stained Protein Marker).

SEC-MALS
CD3E & CD3D MALS images

The purity of Biotinylated Human CD3E&CD3D Heterodimer Protein, Fc,His Tag&Fc,Flag Tag, primary amine labeling (Cat. No. CDD-H82W0) is more than 90% and the molecular weight of this protein is around 80-95 kDa verified by SEC-MALS.

Report
Bioactivity-ELISA
CD3E & CD3D ELISA

Immobilized Biotinylated Human CD3E&CD3D Heterodimer Protein, Fc,His Tag&Fc,Flag Tag, primary amine labeling (Cat. No. CDD-H82W0) at 1 μg/mL (100 μL/well)on streptavidin (Cat. No. STN-N5116) precoated (0.2 μg/well) plate, can bind Monoclonal Anti-Human CD3 Antibody, Mouse IgG2a (Clone: OKT3), premium grade (Cat. No. CDE-M120a) with a linear range of 2-20 ng/mL (QC tested).

Protocol
CD3E & CD3D ELISA

Immobilized Human BCMA Protein, His Tag (HPLC-verified) (Cat. No.BCA-H522y) at 2 μg/mL, add increasing concentrations of Bispecific T cell Engager (CD3 X BCMA) and then add Biotinylated Human CD3E&CD3D Heterodimer Protein, Fc,His Tag&Fc,Flag Tag, primary amine labeling (Cat. No. CDD-H82W0) at 1 μg/mL. Detection was performed using HRP-conjugated streptavidin with sensitivity of 0.39 ng/mL (Intact assay, Routinely tested).

Protocol
Bioactivity-SPR
CD3E & CD3D SPR

Biotinylated Human CD3E&CD3D Heterodimer Protein, Fc,His Tag&Fc,Flag Tag, ultra sensitivity (primary amine labeling) (MALS verified) (Cat. No. CDD-H82W0) captured on Biotin CAP-Series S Sensor Chip can bind Monoclonal Anti-Human CD3 Antibody, Mouse IgG2a (Clone: OKT3) with an affinity constant of 17.9 nM as determined in a SPR assay (Biacore 8K) (Routinely tested).

Protocol
CD3E & CD3D SPR

Biotinylated Human CD3E&CD3D Heterodimer Protein, Fc,His Tag&Fc,Flag Tag, ultra sensitivity (primary amine labeling) (MALS verified) (Cat. No. CDD-H82W0) captured on Biotin CAP-Series S Sensor Chip can bind Bispecific T-cell Engager (CD3 X BCMA) with an affinity constant of 0.45 nM as determined in a SPR assay (Biacore 8K) (Routinely tested).

Protocol
Bioactivity-BLI
CD3E & CD3D BLI

Loaded Biotinylated Human CD3E&CD3D Heterodimer Protein, Fc,His Tag&Fc,Flag Tag, primary amine labeling (Cat. No. CDD-H82W0) on SA Biosensor, can bind Monoclonal Anti-Human CD3 Antibody, Mouse IgG2a (Clone: OKT3), premium grade (Cat. No. CDE-M120a) with an affinity constant of 1.07 nM as determined in BLI assay (ForteBio Octet Red96e) (Routinely tested).

Protocol
CD3E & CD3D BLI

Loaded Biotinylated Human CD3E&CD3D Heterodimer Protein, Fc,His Tag&Fc,Flag Tag, primary amine labeling (Cat. No. CDD-H82W0) on SA Biosensor, can bind UCHT1 with an affinity constant of 21.8 pM as determined in BLI assay (ForteBio Octet Red96e) (Routinely tested).

Protocol
CD3E & CD3D BLI

Loaded Biotinylated Human CD3E&CD3D Heterodimer Protein, Fc,His Tag&Fc,Flag Tag, primary amine labeling (Cat. No. CDD-H82W0) on SA Biosensor, can bind Bispecific BCMA×CD3 T cell-engaging Antibody (Cat. No. CDA-H52Hb) with an affinity constant of 2.05 nM as determined in BLI assay (ForteBio Octet Red96e) (Routinely tested).

Protocol
  • Background
    T-cell surface glycoprotein CD3 delta & CD3 epsilon chain, also known as CD3D & CD3E or CD3D&CD3E respectively, are single-pass type I membrane proteins. CD3D, together with CD3- epsilon(CD3E) , CD3-gamma and CD3-zeta, and the T-cell receptor alpha/beta and gamma/delta heterodimers, forms the T cell receptor-CD3 complex. T cell receptor-CD3 complex plays an important role in coupling antigen recognition to several intracellular signal-transduction pathways.
  • Clinical and Translational Updates

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