Recombinant Proteins iso9001 ISO13485
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Biotinylated Human CD3E&CD3D Heterodimer Protein, Fc,His Tag&Fc,Flag Tag, ultra sensitivity (primary amine labeling) (MALS verified)

分子别名(Synonym)

CD3E & CD3D,CD3 delta & CD3 epsilon,CD3 delta / epsilon

表达区间及表达系统(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)). It is the biotinylated form of Human CD3E&CD3D Heterodimer Protein, Fc,His Tag&Fc,Flag Tag (CDD-H52W0).

Predicted N-terminus: Asp 23 (CD3E) & Phe 22 (CD3D)

蛋白结构(Molecular Characterization)

Online(Asp 23 - Asp 126 (CD3E) & Phe 22 - Ala 105 (CD3D)) NP_000724.1 (CD3E) & NP_000723.1 (CD3D)

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 under reducing (R) condition due to glycosylation.

标记(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.

内毒素(Endotoxin)

Less than 1.0 EU per μg by the LAL method.

纯度(Purity)

>95% as determined by SDS-PAGE.

>90% as determined by SEC-MALS.

制剂(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.

重构方法(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.

存储(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.
 

电泳(SDS-PAGE)

Biotinylated Human CD3E&CD3D Heterodimer Protein, Fc,His Tag&Fc,Flag Tag, primary amine labeling (Cat. No. CDD-H82W0) SDS-PAGE gel

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 overnight with Coomassie Blue. The purity of the protein is greater than 95%.

SEC-MALS

Biotinylated Human CD3E&CD3D Heterodimer Protein, Fc,His Tag&Fc,Flag Tag, primary amine labeling (Cat. No. CDD-H82W0) MALS images

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

Report

 

活性(Bioactivity)-ELISA

Biotinylated Human 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), Ultra-low endotoxin (Cat. No. CDE-M120a) with a linear range of 2-20 ng/mL (QC tested).

Protocol

Biotinylated Human  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

活性(Bioactivity)-SPR

Biotinylated Human 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

Biotinylated Human 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

 

活性(Bioactivity)-BLI

Biotinylated Human 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), Ultra-low endotoxin (Cat. No. CDE-M120a) with an affinity constant of 1.07 nM as determined in BLI assay (ForteBio Octet Red96e) (Routinely tested).

Protocol

Biotinylated Human 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

Biotinylated Human 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

 

背景(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.

 

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