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Biotinylated Human CD3E & CD3D, ultra sensitivity (primary amine labeling)  pdf  pdf  pdf


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Synonym

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

Source

MABSol® Biotinylated Human CD3 epsilon & CD3 delta (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) & AAH39035 (CD3D)). It is the biotinylated form of Human CD3 epsilon & CD3 delta Protein (Cat # CDD-H52W0).

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

Molecular Characterization

CD3E(Asp 23 - Asp 126)NP_000724.1
FC(Pro 100 - Lys 330)P01857
Poly-his
CD3D(Phe 22 - Ala 105)AAH39035
FC(Pro 100 - Lys 330)P01857
Flag

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), 37.5 kDa (CD3D) . The protein migrates as 40-55 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 2.5-4.5 as determined by the HABA assay.

Endotoxin

Less than 1.0 EU per μg by the LAL method.

Purity

>95% as determined by 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 CD3E & CD3D (Cat. No. CDD-H82W0) SDS-PAGE gel

Biotinylated Human CD3E & CD3D 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 CD3E & CD3D (Cat. No. CDD-H82W0) ELISA bioactivity

Immobilized Biotinylated Human CD3E & CD3D (Cat. No. CDD-H82W0) can bind OKT3 with a linear range of 2-39 ng/mL. The biotinylated CD3E & CD3D heterodimers are coated at 1 ug/mL (100 μL/well) on a streptavidin coated plates (QC tested).

Request for protocol

Biotinylated Human CD3E & CD3D (Cat. No. CDD-H82W0) ELISA bioactivity

Immobilized SP34-2 at 5 μg/mL (100 μL/well) can bind Biotinylated Human CD3E & CD3D, His Tag, Fc Tag, Flag Tag (Cat. No. CDD-H82W0) with a linear range of 1-20 ng/mL (Routinely tested).

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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.

References

Please contact us via TechSupport@acrobiosystems.com if you have any question on this product.

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