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Biotinylated Human CD3 epsilon&CD3 delta Heterodimer Protein, His,Avitag™&Tag Free (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)

Biotinylated Human CD3E&CD3D Heterodimer Protein, His,Avitag&Tag Free (CDD-H82W6) is expressed from human 293 cells (HEK293). It contains AA Asp 23 - Asp 126 (CD3E) & Phe 22 - Ala 105 (CD3D) (Accession # P07766-1(CD3E) & P04234-1(CD3D)).

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)) P07766-1 (CD3E) & P04234-1 (CD3D)

Biotinylated Human CD3E&CD3D Heterodimer Protein, His,Avitag&Tag Free is produced by co-expression of CD3E and CD3D, has a calculated MW of 18.6 kDa (CD3E) and 13.4 kDa (CD3D). Subunit CD3E is fused with a polyhistidine tag and an Avi tag (Avitag™) at the C-terminus and subunit CD3D contains no tag. As a result of glycosylation, the heterodimer protein migrates as 20-28 kDa under reducing (R) condition, and 38-47 kDa under non-reducing (NR) condition (SDS-PAGE).

标记(Biotinylation)

Biotinylation of this product is performed using Avitag™ technology. Briefly, the single lysine residue in the Avitag is enzymatically labeled with biotin.

蛋白标记度(Biotin:Protein Ratio)

The biotin to protein ratio is 0.5-1 as determined by the HABA assay.

内毒素(Endotoxin)

Less than 1.0 EU per μg by the LAL method.

纯度(Purity)

>90% as determined by SDS-PAGE.

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, His,Avitag&Tag Free (Cat. No. CDD-H82W6) SDS-PAGE gel

Biotinylated Human CD3E&CD3D Heterodimer Protein, His,Avitag&Tag Free on SDS-PAGE under reducing (R) and non-reducing (NR) conditions. The gel was stained overnight with Coomassie Blue. The purity of the protein is greater than 90%.

SEC-MALS

Biotinylated Human CD3E&CD3D Heterodimer Protein, His,Avitag&Tag Free (Cat. No. CDD-H82W6) MALS images

The purity of Biotinylated Human CD3E&CD3D Heterodimer Protein, His,Avitag&Tag Free (Cat. No. CDD-H82W6) was more than 85% and the molecular weight of this protein is around 40-60 kDa verified by SEC-MALS.

Report

 

活性(Bioactivity)-ELISA

Biotinylated Human ELISA

Immobilized Biotinylated Human CD3E&CD3D Heterodimer Protein, His,Avitag&Tag Free (Cat. No. CDD-H82W6) at 1 μg/mL (100 μL/well) on Streptavidin (Cat. No. STN-N5116) precoated (0.5 μg/well) plate, can bind Monoclonal Anti-Human CD3 Antibody, Mouse IgG2a (Clone: OKT3) (Cat. No. CDE-M120a) with a linear range of 0.2-6 ng/mL (QC tested).

Protocol

Biotinylated Human  ELISA

Immobilized Biotinylated Human CD3E&CD3D Heterodimer Protein, His,Avitag&Tag Free (Cat. No. CDD-H82W6) at 1 μg/mL (100 μL/well) on Streptavidin (Cat. No. STN-N5116) precoated (0.5 μg/well) plate, can bind UCHT1 with a linear range of 0.1-3 ng/mL (Routinely tested).

Protocol

Biotinylated Human ELISA

Quantitative Analysis of Bispecific T-cell Engager (CD3 X BCMA) in human serum.
Immobilized Human BCMA, His Tag (Cat. No. BCA-H522y) at 2 μg/mL, add increasing concentrations of Bispecific T cell Engager (CD3 X BCMA) in 10% human serum and then add Biotinylated Human CD3E&CD3D Heterodimer Protein, His,Avitag&Tag Free (Cat. No. CDD-H82W6) at 0.2 μg/mL. Detection was performed using HRP-conjugated streptavidin with sensitivity of 15 ng/mL (Routinely tested).

Protocol

活性(Bioactivity)-SPR

Biotinylated Human SPR

Biotinylated Human CD3E&CD3D Heterodimer Protein, His,Avitag&Tag Free (Cat. No. CDD-H82W6) captured on Biotin CAP - Series S sensor Chip can bind Anti-Human CD3 Antibody, Mouse IgG2a (Clone: OKT3) with an affinity constant of 22.5 nM as determined in a SPR assay (Biacore T200) (Routinely tested).

Protocol

 

文献引用(Citations)

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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