Recombinant Human CD3 epsilon Fc Chimera Avi-tag Protein, CF

Measured by its binding ability in a functional ELISA. Biotinylated Recombinant Human CD3 epsilon Fc Chimera Avi-tag Protein (Catalog # AVI9850) binds to Human CD3 epsilon Antibody (MAB1067) with an ED50 of 0.600-6.00 ng/mL.

2 μg/lane of Biotinylated Recombinant Human CD3 epsilon Fc Chimera Avi-tag Protein (Catalog # AVI9850) was resolved with SDS-PAGE under reducing (R) and non-reducing (NR) conditions and visualized by Coomassie® Blue staining, showing bands at 45-60 kDa and 90-120 kDa, respectively.

• Reactivity: Hu
• Applications: Bioactivity
• Format: Carrier-Free

Recombinant Human CD3 epsilon Fc Chimera Avi-tag Protein, CF Summary

• Details of Functionality: Measured by its binding ability in a functional ELISA. Biotinylated Recombinant Human CD3 epsilon Fc Chimera Avi-tag binds to Human CD3 epsilon Antibody (Catalog # MAB10670) with an ED₅₀ of 0.600-6.00 ng/mL.
• Source: Human embryonic kidney cell, HEK293-derived human CD3 epsilon protein
  

  Human CD3 epsilon (Asp23-Asp126) Accession # P07766.2	IEGRMD	Human IgG1 (Pro100-Lys330)	Avi-tag
  N-terminus			C-terminus

• N-terminal Sequence: Gln22 inferred from enzymatic pyroglutamate treatment revealing Asp23
• Structure / Form: Disulfide-linked homodimer
  Biotinylated via Avi-tag
• Protein/Peptide Type: Recombinant Proteins
• Purity: >95%, by SDS-PAGE visualized with Silver Staining and quantitative densitometry by Coomassie® Blue Staining
• Endotoxin Note: <0.10 EU per 1 μg of the protein by the LAL method.

Applications/Dilutions

• Dilutions:
  • Bioactivity
• Theoretical MW: 40 kDa.
  Disclaimer note: The observed molecular weight of the protein may vary from the listed predicted molecular weight due to post translational modifications, post translation cleavages, relative charges, and other experimental factors.
• SDS-PAGE: 45-60 kDa, under reducing conditions.

Packaging, Storage & Formulations

• Storage: Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
  • 12 months from date of receipt, -20 to -70 °C as supplied.
  • 1 month, 2 to 8 °C under sterile conditions after reconstitution.
  • 3 months, -20 to -70 °C under sterile conditions after reconstitution.
• Buffer: Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose.
• Purity: >95%, by SDS-PAGE visualized with Silver Staining and quantitative densitometry by Coomassie® Blue Staining
• Reconstitution Instructions: Reconstitute at 250 μg/mL in water.

Notes

This product is produced by and ships from R&D Systems, Inc., a Bio-Techne brand.

Alternate Names for Recombinant Human CD3 epsilon Fc Chimera Avi-tag Protein, CF

• CD3 epsilon
• CD3e antigen
• CD3e antigen, epsilon polypeptide (TiT3 complex)
• CD3e molecule, epsilon (CD3-TCR complex)
• CD3e
• CD3-epsilon
• FLJ18683
• T3E
• T-cell antigen receptor complex, epsilon subunit of T3
• T-cell surface antigen T3/Leu-4 epsilon chain
• T-cell surface glycoprotein CD3 epsilon chain
• TCRE

Background

CD3 epsilon (T-Cell Surface Glycoprotein CD3 Epsilon) is a type I transmembrane protein that belongs to the Ig (Immunoglobulin) superfamily. Human CD3 epsilon consists of 104 amino acid (aa) extracellular domain, 26 aa transmembrane domain and 55 aa cytoplasmic domain (1). With the extracellular domain, human CD3 epsilon shares 44.2% and 49% homology with mouse and rat CD3 epsilon respectively. There are four CD3 proteins: CD3δ, CD3 epsilon , CD3 gamma , and CD3 zeta . All CD3 proteins contain ITAM (Immunoreceptor Tyrosine-based Activation Motifs) in the cytoplasmic tail, which becomes phosphorylated by Src family protein tyrosine kinases LCK and FYN upon TCR engagement (2). CD3 proteins form heterodimers of CD3δ/CD3 epsilon and CD3 gamma /CD3 epsilon , which bind to TCR and form trimeric TCR alpha /CD3 epsilon /CD3 gamma and TCR beta /CD3 gamma /CD3 epsilon . The resulting heterohexamer further associates with CD3 zeta homodimer and forms TCR/CD3 signaling complex (3, 4). Similar complex is also formed with TCR gamma and TCRδ (5). CD3 epsilon plays essential role in adaptive immune response. Mutations in CD3 epsilon lead to SCID (severe combined immunodeficiency) (6). Our Avi-tag Biotinylated CD3 epsilon  features biotinylation at a single site contained within the Avi-tag, a unique 15 amino acid peptide. Protein orientation will be uniform when bound to streptavidin-coated surface due to the precise control of biotinylation and the rest of the protein is unchanged so there is no interference in the protein's bioactivity.

1. Gold, D.P. et al. (1986) Nature 321:431.
2. Barber, E.K. et al. (1989) Proc. Nat. Acad. Sci. U.S.A. 86:3277.
3. Manolios, N. et al. (1991) EMBO J 10:1643.
4. Dietrich, J. et al. (1999) Eur J Immunol 29:1719.
5. Pan, Q. et al. (2006) Mol Immunol 43:1741.
6. Soudais, C. et al. (1993) Nat. Genet. 3:77.