Recombinant Human CD277/BTN3A1 Fc Avi-tag Protein, CF

Measured by its binding ability in a functional ELISA. Biotinylated Recombinant Human CD277/BTN3A1 Fc Chimera Avi-tag Protein (Catalog # AVI8539) binds to Human BTN3A1/2/3 Antibody (MAB7136) with an ED50 of 0.500‑5.00 ng/mL.

2 μg/lane of Biotinylated Recombinant Human CD277/BTN3A1 Fc Chimera Avi-tag Protein (Catalog # AVI8539) was resolved with SDS-PAGE under reducing (R) and non-reducing (NR) conditions and visualized by Coomassie® Blue staining, showing bands at 57‑65 kDa and 110‑130 kDa, respectively.

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

Recombinant Human CD277/BTN3A1 Fc Avi-tag Protein, CF Summary

• Additional Information: Fc Chimera
• Details of Functionality: Measured by its binding ability in a functional ELISA. Biotinylated Recombinant Human CD277/BTN3A1 Fc Chimera Avi-tag binds to Human BTN3A1/2/3 Antibody (Catalog # MAB7136) with an ED₅₀ of 0.500-5.00 ng/mL.
• Source: Chinese Hamster Ovary cell line, CHO-derived human CD277/BTN3A1 protein
  

  Human CD277/BTN3A1 (Gln30-Gly254) Accession # O00481.3	IEGRMD	Human IgG1 (Pro100-Lys330)	Avi-tag
  N-terminus			C-terminus

• N-terminal Sequence: Gln30 inferred from enzymatic pyroglutamate treatment revealing Phe31
• 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
• Purity Statement: Antigen Affinity-purified
• Endotoxin Note: <0.10 EU per 1 μg of the protein by the LAL method.

Applications/Dilutions

• Dilutions:
  • Bioactivity
• Theoretical MW: 53 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: 57-65 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 500 μg/mL in PBS.

Notes

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

Alternate Names for Recombinant Human CD277/BTN3A1 Fc Avi-tag Protein, CF

• BT3.1
• BTF5
• BTN3A1
• butyrophilin, subfamily 3, member A1
• CD277
• dJ45P21.3 (butyrophilin, subfamily 3, member A1)

Background

Butyrophilin 3A1 (also called BTN3A1) is a 57 kDa type I transmembrane glycoprotein member of the Ig superfamily. It is expressed on a wide variety of immune cells. Similar to BTN3A2 and BTN3A3, BTN3A1 (484 amino acids) is composed of an extracellular N-terminal IgV and a membrane-proximal IgC domain followed by a transmembrane domain and a cytoplasmic tail. These Ig domains are also found in B7 family co-stimulatory molecules, suggesting structural and functional similarities between the two protein families (1). The intracellular portion of BTN3A1 contains a B30.2 domain (2). Although the B30.2 domain of BTN1A1 binds to xanthine oxidoreductase (XOR) and is conserved among BTN1A1 orthologs, this interaction with XOR is not shared by BTN3A1 (3). The B30.2 domain of butyrophilins also functions as a sensor for detecting changes in intracellular phopho-antigen (pAg) concentrations produced during tumorigenesis and microbial infections (4, 5). The specific binding of pAg by the B30.2 domain of BTN3A1 induces a conformational change in its ECD, leading to the activation of V gamma 9Vδ2 T cells (6). Thus, BTN3A1 acts as a critical protein for the activation of V gamma 9Vδ2 T cells following detection of distressed cells (7). The anti-tumor responses of V gamma 9Vδ2 T cells may be enhanced with agonistic anti-BTNA3 antibodies (8). No BTN3A1 homolog has yet been identified in rodents. However, Human BTN3A1 shares 92.4% and 91.6% sequence identity with baboon and rhesus monkey BTN3A1 respectively (9).  Our Avi-tag Biotinylated  human BTN3A1 Fc chimera 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. Abeler-Dorner, L. et al. (2012) Trends Immunol. 33:34.
2. Rhodes, D.A. et al. (2001) Genomics 71:351.
3. Jeong, J. et al. (2009) J. Biol. Chem. 284:22444.
4. Harly, C. et al. (2012) Blood 120:2269.
5. Constant, P. et al. (1994) Science 264:267.
6. Sandstrom, A. et al. (2014) Immunity 40:490.
7. Harly, C. et al. (2015) Front. Immunol. 5:657.
8. Bonneville, M. et al. (2006) Curr. Opin. Immunol. 18:539.
9. Wang, H. et al. (2013) J.Immunol. 191:1029.