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    R718 Mouse Anti-MR1
    R718 Mouse Anti-MR1

    Flow cytometric analysis of MR1 expression on viable Human peripheral blood lymphocytes.  Human peripheral blood mononuclear cells (PBMCs) were either not cultured with Acetyl-6-Formylpterin (No Ac-6-FP; 2.3 µg/ml (10 µM); Cayman Chemicals Cat. No. 23303; Left Plot) or cultured (16 h) with Ac-6-FP (+ Ac-6-FP; Right Plot) as indicated. The cells were then preincubated with Human BD Fc Block™ (Cat. No. 564219/564220) and stained with either BD Horizon™ R718 Mouse IgG1, κ Isotype Control (Cat. No. 566928; dashed line histograms) or BD Horizon™ R718 Mouse anti-MR1 antibody (Cat. No. 568420/568421; solid line histograms) at 1 µg/test. DAPI (4',6-Diamidino-2-Phenylindole, Dihydrochloride) 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) lymphocytes. 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.

    R718 Mouse Anti-MR1

    Flow cytometric analysis of MR1 expression on viable B16-F0 cells. Cells from the Mouse B16-F0 (Skin Melanoma, ATCC® CRL-6322™) cell line were either not cultured with Acetyl-6-Formylpterin (No Ac-6-FP; 2.3 µg/ml (10 µM); Cayman Chemicals Cat. No. 23303; Left Plot) or cultured (16 h) with Ac-6-FP (+ Ac-6-FP; Right Plot) as indicated. The cells were then preincubated with Purified Rat Anti-Mouse CD16/CD32 antibody (Mouse BD Fc Block™) [Cat. No. 553141/553142] and stained with either BD Horizon™ R718 Mouse IgG1, κ Isotype Control (Cat. No. 566928; dashed line histograms) or BD Horizon™ R718 Mouse Anti-MR1 antibody (Cat. No. 568420/568421; solid line histograms) at 1 µg/test. DAPI (4',6-Diamidino-2-Phenylindole, Dihydrochloride) Solution 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.

    R718 Mouse Anti-MR1 R718 Mouse Anti-MR1

    Flow cytometric analysis of MR1 expression on viable Human peripheral blood lymphocytes.  Human peripheral blood mononuclear cells (PBMCs) were either not cultured with Acetyl-6-Formylpterin (No Ac-6-FP; 2.3 µg/ml (10 µM); Cayman Chemicals Cat. No. 23303; Left Plot) or cultured (16 h) with Ac-6-FP (+ Ac-6-FP; Right Plot) as indicated. The cells were then preincubated with Human BD Fc Block™ (Cat. No. 564219/564220) and stained with either BD Horizon™ R718 Mouse IgG1, κ Isotype Control (Cat. No. 566928; dashed line histograms) or BD Horizon™ R718 Mouse anti-MR1 antibody (Cat. No. 568420/568421; solid line histograms) at 1 µg/test. DAPI (4',6-Diamidino-2-Phenylindole, Dihydrochloride) 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) lymphocytes. 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.

    Flow cytometric analysis of MR1 expression on viable B16-F0 cells. Cells from the Mouse B16-F0 (Skin Melanoma, ATCC® CRL-6322™) cell line were either not cultured with Acetyl-6-Formylpterin (No Ac-6-FP; 2.3 µg/ml (10 µM); Cayman Chemicals Cat. No. 23303; Left Plot) or cultured (16 h) with Ac-6-FP (+ Ac-6-FP; Right Plot) as indicated. The cells were then preincubated with Purified Rat Anti-Mouse CD16/CD32 antibody (Mouse BD Fc Block™) [Cat. No. 553141/553142] and stained with either BD Horizon™ R718 Mouse IgG1, κ Isotype Control (Cat. No. 566928; dashed line histograms) or BD Horizon™ R718 Mouse Anti-MR1 antibody (Cat. No. 568420/568421; solid line histograms) at 1 µg/test. DAPI (4',6-Diamidino-2-Phenylindole, Dihydrochloride) Solution 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.

    Product Details
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    BD Horizon™
    HLALS; MHC class I-like antigen MR-1; MHC class-I related-gene protein
    Human (QC Testing), Mouse (Tested in Development), Rat (Reported)
    Mouse IgG1, κ
    MR1 Extracellular Domain Complexed with β2m
    Flow cytometry (Routinely Tested)
    0.2 mg/ml
    3140, 15064
    AB_3684259
    Aqueous buffered solution containing ≤0.09% sodium azide.
    RUO


    Preparation And Storage

    Store undiluted at 4°C and protected from prolonged exposure to light. Do not freeze. The monoclonal antibody was purified from tissue culture supernatant or ascites by affinity chromatography. The antibody was conjugated to the dye under optimum conditions and unreacted dye was removed.
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    Recommended Assay Procedures

    BD® CompBeads can be used as surrogates to assess fluorescence spillover (compensation). When fluorochrome conjugated antibodies are bound to BD® CompBeads, they have spectral properties very similar to cells. However, for some fluorochromes there can be small differences in spectral emissions compared to cells, resulting in spillover values that differ when compared to biological controls. It is strongly recommended that when using a reagent for the first time, users compare the spillover on cells and BD® CompBeads to ensure that BD® CompBeads are appropriate for your specific cellular application.

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    Product Notices

    1. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
    2. Since applications vary, each investigator should titrate the reagent to obtain optimal results.
    3. An isotype control should be used at the same concentration as the antibody of interest.
    4. 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.
    5. This product is provided under an Agreement between BIOTIUM and BD Biosciences. This product, and only in the amount purchased by buyer, may be used solely for buyer’s own internal research, in a manner consistent with the accompanying product literature. No other right to use, sell or otherwise transfer (a) this product, or (b) its components is hereby granted expressly, by implication or by estoppel. This product is for research use only. Diagnostic uses require a separate license from Biotium, Inc. For information on purchasing a license to this product including for purposes other than research, contact Biotium, Inc., 3159 Corporate Place, Hayward, CA 94545, Tel: (510) 265-1027. Fax: (510) 265-1352. Email: btinfo@biotium.com.
    6. Human donor specific background has been observed in relation to the presence of anti-polyethylene glycol (PEG) antibodies, developed as a result of certain vaccines containing PEG, including some COVID-19 vaccines. We recommend use of BD Horizon Brilliant™ Stain Buffer in your experiments to help mitigate potential background. For more information visit https://www.bdbiosciences.com/en-us/support/product-notices.
    7. Species cross-reactivity detected in product development may not have been confirmed on every format and/or application.
    8. Please refer to http://regdocs.bd.com to access safety data sheets (SDS).
    9. Alexa Fluor™ is a trademark of Life Technologies Corporation.
    10. For U.S. patents that may apply, see bd.com/patents.
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    568420 Rev. 2
    Antibody Details
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    8F2.F9

    The 8F2.F9 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.

    568420 Rev. 2
    Format Details
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    R718
    The BD Horizon™ Red 718 (R718) Dye is part of the BD red family of dyes. It is a small organic fluorochrome with an excitation maximum (Ex Max) at 695-nm and an emission maximum (Em Max) at 718-nm. Driven by BD innovation, R718 is designed to be excited by the red laser (627–640-nm) and detected using an optical filter centered near 720-nm (e.g., a 720/40-nm bandpass filter). R718 is a brighter alternative to Alexa Fluor™ 700. R718 is also a bright small molecule alternative to APC-R700 with lower spread into the APC detector. Please ensure that your instrument’s configurations (lasers and optical filters) are appropriate for this dye.
    altImg
    R718
    Red 627-640 nm
    695 nm
    718 nm
    568420 Rev.2
    Citations & References
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    View product citations for antibody "568420" on CiteAb

    Development References (10)

    1. 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. (Biology: Flow cytometry). View Reference
    2. 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
    3. 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
    4. 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
    5. Lamichhane R, Ussher JE. Expression and trafficking of MR1.. Immunology. 2017; 151(3):270-279. (Biology). View Reference
    6. 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
    7. 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
    8. 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
    9. Walter L, Günther E. Isolation and molecular characterization of the rat MR1 homologue, a non-MHC-linked class I-related gene.. Immunogenetics. 1998; 47(6):477-82. (Biology). View Reference
    10. 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
    View All (10) View Less
    568420 Rev. 2

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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.