Mouse Anti-BTN3A1 Recombinant Antibody (clone 20.1) (CAT#: PABL-414)

Recombinant Mouse Antibody (20.1) is capable of binding to BTN3A1. This agonist antibody, 20.1, binds to the extracellular domain of BTN3A molecules and induces Vγ9Vδ2 activation with similar kinetics and potency as with addition of PiPs.


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ELISA

Figure 1 HLA haplo-identical HeLa-L and HeLa-M cells vary in their ability to present phosphoantigens to Vγ9Vδ2 T cells.

Figure 1 HLA haplo-identical HeLa-L and HeLa-M cells vary in their ability to present phosphoantigens to Vγ9Vδ2 T cells.

Activation of Vγ9/Vδ2 T cells detected by ELISA for IFN-γ using a concentration gradient of HMB-PP, agonist antibody CD277 20.1 and T cells expanded from two separate donors. Activation was induced by pretreatment of HeLa cells using anti-BTN3A monoclonal antibody 20.1 for 30 min. Graphs show mean and SD from n = 2 and are representative of two independent measurements.

Rhodes, D. A., Chen, H. C., Williamson, J. C., Hill, A., Yuan, J., Smith, S., ... & Eberl, M. (2018). regulation of human γδ T cells by BTn3a1 Protein stability and aTP-Binding cassette Transporters. Frontiers in immunology, 9, 662.

FC

Figure 2 Transcripts for the three BTN3A isoforms are unchanged in HeLa-L and HeLa-M.

Figure 2 Transcripts for the three BTN3A isoforms are unchanged in HeLa-L and HeLa-M.

Analysis of activation markers MHC-I (w6/32), MHC-II (L243), and BTN3A (CD277 20.1) by flow cytometry in HeLa cells treated with or without IFN-γ. Results representative of triplicate experiments. Red lines indicate IFN-γ treated and black lines untreated samples. Solid lines are HeLa-M and dotted lines HeLa-L.

Rhodes, D. A., Chen, H. C., Williamson, J. C., Hill, A., Yuan, J., Smith, S., ... & Eberl, M. (2018). regulation of human γδ T cells by BTn3a1 Protein stability and aTP-Binding cassette Transporters. Frontiers in immunology, 9, 662.

WB

Figure 3 Western blot of HeLa-L cells transiently transfected with expression vectors for BTN3A1, BTN3A2, and BTN3A3.

Figure 3 Western blot of HeLa-L cells transiently transfected with expression vectors for BTN3A1, BTN3A2, and BTN3A3.

Monoclonal anti-BTN3A (CD277 20.1) antibody and polyclonal sera 056 and B6, directed to B30.2 domains of BTN3A1 and BTN3A3, respectively, were used. Anti-GFP and calnexin (CNX) antibodies provided loading controls for expression of transgene and endogenous proteins, respectively.

Rhodes, D. A., Chen, H. C., Williamson, J. C., Hill, A., Yuan, J., Smith, S., ... & Eberl, M. (2018). regulation of human γδ T cells by BTn3a1 Protein stability and aTP-Binding cassette Transporters. Frontiers in immunology, 9, 662.

FuncS

Figure 4 CD107a expression on the human Vγ9Vδ2 T cell line GUI following incubation with either 20.1 full-lengthantibody (open circles), 20.1 Fab fragments (filled triangles), or 20.1 scFv (filled circles).

Figure 4 CD107a expression on the human Vγ9Vδ2 T cell line GUI following incubation with either 20.1 full-lengthantibody (open circles), 20.1 Fab fragments (filled triangles), or 20.1 scFv (filled circles).

The data are presented as thepercentages of CD107a+ γδT cells and are representative of more than three experiments.

Palakodeti, A., Sandstrom, A., Sundaresan, L., Harly, C., Nedellec, S., Olive, D., ... & Adams, E. J. (2013). The Molecular Basis for Modulation of Human V;# 947; 9V [delta] 2 T Cell Responses by CD277/Butyrophilin-3 (BTN3A)-specific Antibodies. J. Biol. Chem., 287(39); 09, 2012).


Specifications

  • Immunogen
  • Human butyrophilin, subfamily 3, member A1
  • Host Species
  • Mouse
  • Derivation
  • Mouse
  • Type
  • Mouse IgG
  • Specificity
  • BTN3A1
  • Species Reactivity
  • Human
  • Clone
  • 20.1
  • Applications
  • ELISA, FC, WB, FuncS

Product Property

  • Purity
  • >95% as determined by analysis by SDS-PAGE
  • Concentration
  • Please refer to the vial label for the specific concentration.
  • Buffer
  • PBS
  • Preservative
  • No preservatives
  • Storage
  • Centrifuge briefly prior to opening vial. Store at +4°C short term (1-2 weeks). Aliquot and store at -20°C long term. Avoid repeated freeze/thaw cycles.

Applications

  • Application Notes
  • This antibody has been reported for use in Enzyme-Linked Immunosorbent Assay, Flow Cytometry, Western Blot and Function Assay.

Target

  • Alternative Names
  • BTN3A1; butyrophilin, subfamily 3, member A1; BTF5; BT3.1; CD277; BTN3.1; butyrophilin subfamily 3 member A1; dJ45P21.3 (butyrophilin, subfamily 3, member A1)

Product Notes

This is a product of Creative Biolabs' Hi-Affi™ recombinant antibody portfolio, which has several benefits including:

• Increased sensitivity
• Confirmed specificity
• High repeatability
• Excellent batch-to-batch consistency
• Sustainable supply
• Animal-free production

See more details about Hi-Affi™ recombinant antibody benefits.

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For Research Use Only. Not For Clinical Use.

For research use only. Not intended for any clinical use. No products from Creative Biolabs may be resold, modified for resale or used to manufacture commercial products without prior written approval from Creative Biolabs.

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