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Flow cytometric analysis of MR1 expression on Human peripheral blood lymphocytes. Human peripheral blood mononuclear cells (PBMCs) were either not cultured with Acetyl-6-Formylpterin (No Ac-6-FP; Left Plot) or cultured (16 h) with Ac-6-FP (+ Ac-6-FP; 2.3 µg/ml (10 µM); Cayman Chemicals Cat. No. 23303; Right Plot) as indicated. The cells were then preincubated with Human BD Fc Block™ (Cat. No. 564219/564220) and stained with either Purified Mouse IgG2a, κ Isotype Control (Cat. No. 553454; dashed line histograms) or Purified Mouse Anti-MR1 antibody (Cat. No. 569050; solid line histograms) at 0.5 µg/test. The cells were secondarily stained with PE Goat Anti-Mouse Ig (Multiple Adsorption) antibody [Cat. No. 550589]. DAPI Solution (Cat. No. 564907) was added to cells right before analysis. The fluorescence histograms showing MR1 expression (or Ig Isotype control staining) were derived from gated events with the forward and side light-scatter characteristics of viable (DAPI-negative) cells. Flow cytometry and data analysis were performed using a BD LSRFortessa™ X-20 Cell Analyzer System and FlowJo™ software. Data shown on this Technical Data Sheet are not lot specific.
BD Pharmingen™ Purified Mouse Anti-MR1
Regulatory Status Legend
Any use of products other than the permitted use without the express written authorization of Becton, Dickinson and Company is strictly prohibited.
Preparation And Storage
Product Notices
- An isotype control should be used at the same concentration as the antibody of interest.
- Caution: Sodium azide yields highly toxic hydrazoic acid under acidic conditions. Dilute azide compounds in running water before discarding to avoid accumulation of potentially explosive deposits in plumbing.
- For U.S. patents that may apply, see bd.com/patents.
- Please refer to http://regdocs.bd.com to access safety data sheets (SDS).
- Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
- Since applications vary, each investigator should titrate the reagent to obtain optimal results.
- Sodium azide is a reversible inhibitor of oxidative metabolism; therefore, antibody preparations containing this preservative agent must not be used in cell cultures nor injected into animals. Sodium azide may be removed by washing stained cells or plate-bound antibody or dialyzing soluble antibody in sodium azide-free buffer. Since endotoxin may also affect the results of functional studies, we recommend the NA/LE (No Azide/Low Endotoxin) antibody format, if available, for in vitro and in vivo use.
- Species cross-reactivity detected in product development may not have been confirmed on every format and/or application.
Companion Products
The 26.5 monoclonal antibody specifically recognizes MHC-related protein 1 (MR1). MRI is a nonclassical MHC class Ib molecule. It is comprised of a ~40 kDa, highly conserved transmembrane a heavy chain that is a type I glycoprotein which is noncovalently-associated with an invariant ß2-microglobulin (ß2m) light chain. The N-terminal extracellular region of the HLA class I heavy chain is comprised of three domains (a1, a2, and a3). The a1 and a2 domains form a closed antigen-binding groove that accommodates small antigens whereas the a3 domain interacts with ß2m. MR1 is an antigen-presenting molecule that is specialized in displaying metabolites derived from microbial riboflavin biosynthesis to a small population of aß T-cells expressing an invariant TCR a chain called mucosal-associated invariant T-cells (MAIT). This function is essential to the development and expansion of MAIT cells. The 8F2.F9 and 26.5 monoclonal antibodies reportedly bind to different MRI epitopes. Clone 26.5 has been shown to crossreact with mouse and rat as well as human MR1.
Development References (10)
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Abós B, Gómez Del Moral M, Gozalbo-López B, López-Relaño J, Viana V, Martínez-Naves E. Human MR1 expression on the cell surface is acid sensitive, proteasome independent and increases after culturing at 26°C.. Biochem Biophys Res Commun. 2011; 411(3):632-6. (Clone-specific: Flow cytometry). View Reference
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Chua WJ, Kim S, Myers N, et al. Endogenous MHC-related protein 1 is transiently expressed on the plasma membrane in a conformation that activates mucosal-associated invariant T cells. J Immunol. 2011; 186(8):4744-4750. (Immunogen). View Reference
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Corbett AJ, Awad W, Wang H, Chen Z. Antigen Recognition by MR1-Reactive T Cells; MAIT Cells, Metabolites, and Remaining Mysteries.. Front Immunol. 2020; 11:1961. (Biology). View Reference
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Huang S, Gilfillan S, Cella M, et al. Evidence for MR1 antigen presentation to mucosal-associated invariant T cells.. J Biol Chem. 2005; 280(22):21183-93. (Immunogen: Flow cytometry). View Reference
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Huang S, Martin E, Kim S, et al. MR1 antigen presentation to mucosal-associated invariant T cells was highly conserved in evolution.. Proc Natl Acad Sci U S A. 2009; 106(20):8290-5. (Clone-specific: Flow cytometry). View Reference
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Lamichhane R, Ussher JE. Expression and trafficking of MR1.. Immunology. 2017; 151(3):270-279. (Biology). View Reference
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McWilliam HE, Eckle SB, Theodossis A, et al. The intracellular pathway for the presentation of vitamin B-related antigens by the antigen-presenting molecule MR1.. Nat Immunol. 2016; 17(5):531-7. (Clone-specific: Flow cytometry). View Reference
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Miley MJ, Truscott SM, Yu YY, et al. Biochemical features of the MHC-related protein 1 consistent with an immunological function.. J Immunol. 2003; 170(12):6090-8. (Biology: Blocking, Flow cytometry, In vivo exacerbation). View Reference
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Salio M, Awad W, Veerapen N, et al. Ligand-dependent downregulation of MR1 cell surface expression.. Proc Natl Acad Sci U S A. 2020; 117(19):10465-10475. (Clone-specific: Flow cytometry). View Reference
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Yan J, Allen S, McDonald E, et al. MAIT Cells Promote Tumor Initiation, Growth, and Metastases via Tumor MR1.. Cancer Discov. 2020; 10(1):124-141. (Clone-specific). View Reference
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Global - Refer to manufacturer's instructions for use and related User Manuals and Technical data sheets before using this products as described
Comparisons, where applicable, are made against older BD Technology, manual methods or are general performance claims. Comparisons are not made against non-BD technologies, unless otherwise noted.
For Research Use Only. Not for use in diagnostic or therapeutic procedures.