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.
Published Data
Gene Expression
ELISA
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.
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.
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).
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).
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Subcellular Location
Figure 1 IF staining of human cell line U-251 MG
Immunofluorescent staining of human cell line U-251 MG shows localization to vesicles.
* Image credit: Image credit: Human Protein Atlas v21.proteinatlas.org/images/12565/if_selected.jpg
Normal Tissue
Figure 2 IHC staining of human kidney
Immunohistochemical staining of human kidney shows strong cytoplasmic positivity in a granular pattern in renal tubules.
* Image credit: Image credit: Human Protein Atlas v21.proteinatlas.org/images/12565/ihc_selected.jpg
Normal Tissue
Figure 3 Cerebral cortex
Endothelial cells
Staining: Medium
Intensity: Moderate
Quantity: 75%-25%
Location: Cytoplasmic/membranous
* Image credit: Image credit: Human Protein Atlas v21.proteinatlas.org/images/12565/40150_B_7_5.jpg
Normal Tissue
Figure 4 Colon
Endothelial cells
Staining: Medium
Intensity: Moderate
Quantity: 75%-25%
Location: Cytoplasmic/membranous
Glandular cells
Staining: High
Intensity: Strong
Quantity:>75%
Location: Cytoplasmic/membranous
Peripheral nerve/ganglion
Staining: Medium
Intensity: Moderate
Quantity: 75%-25%
Location: Cytoplasmic/membranous
* Image credit: Image credit: Human Protein Atlas v21.proteinatlas.org/images/12565/40150_A_9_3.jpg
Normal Tissue
Figure 5 Liver
Hepatocytes
Staining: Medium
Intensity: Moderate
Quantity:>75%
Location: Cytoplasmic/membranous
* Image credit: Image credit: Human Protein Atlas v21.proteinatlas.org/images/12565/40150_A_9_4.jpg
Normal Tissue
Figure 6 Kidney
Cells in glomeruli
Staining: Medium
Intensity: Moderate
Quantity: 75%-25%
Location: Cytoplasmic/membranous
Cells in tubules
Staining: High
Intensity: Strong
Quantity: 75%-25%
Location: Cytoplasmic/membranous
* Image credit: Image credit: Human Protein Atlas v21.proteinatlas.org/images/12565/40150_A_9_5.jpg
Normal Tissue
Figure 7 Testis
Leydig cells
Staining: High
Intensity: Strong
Quantity:>75%
Sertoli cells
Staining: Medium
Intensity: Moderate
Quantity: 75%-25%
* Image credit: Image credit: Human Protein Atlas v21.proteinatlas.org/images/12565/40150_A_6_6.jpg
Normal Tissue
Figure 8 Lymph node
Germinal center cells
Staining: Medium
Intensity: Strong
Quantity: <25%
Location: Cytoplasmic/membranous
Non-germinal center cells
Staining: High
Intensity: Strong
Quantity: 75%-25%
Location: Cytoplasmic/membranous
* Image credit: Image credit: Human Protein Atlas v21.proteinatlas.org/images/12565/40150_A_9_8.jpg
RNA Expression
Figure 9 RNA cell line category: Low cell line specificity
Cell lines ordered by descending RNA expression order.
* Image credit: Image credit: Human Protein Atlas v21.proteinatlas.org/ENSG00000026950-BTN3A1
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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
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Customer Review
There are currently no Customer reviews or questions for PABL-414. Click the button above to contact us or submit your feedback about this product.
Michael Roberts09.Apr,23
Excellent for BTN3A1 Detection
The Anti-BTN3A1 Recombinant Antibody (clone 20-1) has been excellent for our BTN3A1 studies. Its specificity and sensitivity are top-notch, providing clear and reliable results. This antibody has significantly improved the accuracy of our experiments and saved us a lot of troubleshooting time.
Sarah Green11.Jul,22
Effective in Immunoassays
We incorporated this antibody into our immunoassays, and it has been a great asset. The specificity for BTN3A1 is precise, allowing for accurate detection and analysis. The reliable performance has made it an invaluable tool in our lab.
Laura Young25.Sep,21
Consistent High-Quality Results
This antibody has proven to be consistent and reliable in our assays. The high-quality, reproducible results have significantly improved our experiments, making it a crucial part of our research toolkit. The strong affinity and specificity for BTN3A1 have been particularly beneficial.
Q&As
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Is the anti-BTN3A1 recombinant antibody clone 20.1 suitable for use in Western blotting?
A: Yes, the anti-BTN3A1 recombinant antibody clone 20.1 (PABL-414) is suitable for use in Western blotting. It provides specific binding to BTN3A1, allowing for reliable detection in Western blot assays.
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What are the recommended storage conditions for the anti-BTN3A1 recombinant antibody clone 20.1?
A: The recommended storage condition for the anti-BTN3A1 recombinant antibody clone 20.1 (PABL-414) is at -20°C or lower. For short-term storage, it can be kept at 2-8°C. To ensure stability, avoid repeated freeze-thaw cycles.
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Can the anti-BTN3A1 recombinant antibody clone 20.1 be used in immunoprecipitation assays?
A: Yes, the anti-BTN3A1 recombinant antibody clone 20.1 (PABL-414) can be used in immunoprecipitation assays. It provides specific binding to BTN3A1, enabling the successful precipitation of the target protein from complex mixtures.
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Is the anti-BTN3A1 recombinant antibody clone 20.1 effective in ELISA applications?
A: Yes, the anti-BTN3A1 recombinant antibody clone 20.1 (PABL-414) is effective in ELISA applications. It has been validated for use in such assays and provides reliable detection of BTN3A1.
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What is the optimal dilution for using the anti-BTN3A1 recombinant antibody clone 20.1 in immunofluorescence?
A: The optimal dilution for using the anti-BTN3A1 recombinant antibody clone 20.1 (PABL-414) in immunofluorescence is typically 1:100 to 1:500. It is advisable to perform a dilution series to determine the best working concentration for your specific experimental conditions.
View the frequently asked questions answered by Creative Biolabs Support.
Citations
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Girard, Pauline, et al. "Dysfunctional BTN3A together with deregulated immune checkpoints and type I/II IFN dictate defective interplay between pDCs and γδ T cells in melanoma patients, which impacts clinical outcomes." Clinical & Translational Immunology 10.11 (2021): e1329. https://doi.org/10.1002/cti2.1329This study investigates the dysfunctional BTN3A and its interaction with deregulated immune checkpoints and type I/II interferons in the context of melanoma. The research focuses on the roles of plasmacytoid dendritic cells (pDCs) and gamma-delta (γδ) T cells, examining their phenotypic and functional interactions in both the blood and tumor microenvironment of melanoma patients. The study highlights severe alterations in the bidirectional cross-talk between pDCs and γδ T cells in melanoma, noting that while healthy pDCs can drive potent activation of γδ T cells, those from melanoma patients exhibit impaired functionality.
Creative Biolabs provided critical reagents for this study, specifically anti-BTN3A agonist antibody (clone 20.1, Cat# PABL-414) and anti-BTN3A blocking antibody (clone 103.2, Cat# PABL-415). These antibodies were used to assess the functionality of BTN3A on γδ T cells and pDCs within PBMCs and tumor infiltrates. The use of these antibodies was essential in revealing the potential functional impairment of BTN3A in melanoma patients, thereby providing insights into the therapeutic targeting of BTN3A to improve immune responses in cancer therapy.
Cite This Product
To accurately reference this product in your publication, please use the following citation information:
(Creative Biolabs Cat# PABL-414, RRID: AB_3111654)
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Downloadable Resources
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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.
Datasheet
MSDS
COA
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| TAB-1312CL | Mouse Anti-BTN3A1 Recombinant Antibody (TAB-1312CL) | FC, FuncS | Mouse IgG |
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