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      BV421 Mouse Anti-MR1
      BV421 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™ BV421 Mouse IgG1, κ Isotype Control (Cat. No. 562438; dashed line histograms) or BD Horizon™ BV421 Mouse anti-MR1 antibody (Cat. No. 568418/568419; solid line histograms) at 1 µg/test. BD Via-Probe™ Cell Viability 7-AAD Solution (Cat. No. 555815/555816) 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 (7-AAD-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.

      BV421 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™ BV421 Mouse IgG1, κ Isotype Control (Cat. No. 562438; dashed line histograms) or BD Horizon™ BV421 Mouse Anti-MR1 antibody (Cat. No. 568418/568419; solid line histograms) at 1 µg/test. BD Via-Probe™ Cell Viability 7-AAD 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 (7-AAD-negative) cells. Flow cytometry and data analysis were performed using a BD LSRFortessa™ X20 Cell Analyzer System and FlowJo™ software. Data shown on this Technical Data Sheet are not lot specific.

      BV421 Mouse Anti-MR1 BV421 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™ BV421 Mouse IgG1, κ Isotype Control (Cat. No. 562438; dashed line histograms) or BD Horizon™ BV421 Mouse anti-MR1 antibody (Cat. No. 568418/568419; solid line histograms) at 1 µg/test. BD Via-Probe™ Cell Viability 7-AAD Solution (Cat. No. 555815/555816) 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 (7-AAD-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™ BV421 Mouse IgG1, κ Isotype Control (Cat. No. 562438; dashed line histograms) or BD Horizon™ BV421 Mouse Anti-MR1 antibody (Cat. No. 568418/568419; solid line histograms) at 1 µg/test. BD Via-Probe™ Cell Viability 7-AAD 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 (7-AAD-negative) cells. Flow cytometry and data analysis were performed using a BD LSRFortessa™ X20 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_3684257
      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 unconjugated antibody and free dye were 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.

         For optimal and reproducible results, BD Horizon Brilliant Stain Buffer should be used anytime BD Horizon Brilliant dyes are used in a multicolor flow cytometry panel.  Fluorescent dye interactions may cause staining artifacts which may affect data interpretation.  The BD Horizon Brilliant Stain Buffer was designed to minimize these interactions. When BD Horizon Brilliant Stain Buffer is used in in the multicolor panel, it should also be used in the corresponding compensation controls for all dyes to achieve the most accurate compensation. For the most accurate compensation, compensation controls created with either cells or beads should be exposed to BD Horizon Brilliant Stain Buffer for the same length of time as the corresponding multicolor panel. More information can be found in the Technical Data Sheet of the BD Horizon Brilliant Stain Buffer (Cat. No. 563794/566349) or the BD Horizon Brilliant Stain Buffer Plus (Cat. No. 566385).

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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. For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
      6. BD Horizon Brilliant Violet 421 is covered by one or more of the following US patents: 8,158,444; 8,362,193; 8,575,303; 8,354,239.
      7. BD Horizon Brilliant Stain Buffer is covered by one or more of the following US patents: 8,110,673; 8,158,444; 8,575,303; 8,354,239.
      8. 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.
      9. Species cross-reactivity detected in product development may not have been confirmed on every format and/or application.
      10. Please refer to http://regdocs.bd.com to access safety data sheets (SDS).
      11. For U.S. patents that may apply, see bd.com/patents.
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      568418 Rev. 1
      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.

      568418 Rev. 1
      Format Details
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      BV421
      The BD Horizon Brilliant Violet™ 421 (BV421) Dye is part of the BD Horizon Brilliant Violet™ family of dyes. This polymer-technology based dye has an excitation maximum (Ex Max) of 407-nm and an emission maximum (Em Max) at 423-nm. Driven by BD innovation, BV421 is designed to be excited by the violet laser (405-nm) and detected using an optical filter centered near 420-nm (e.g., a 431/28-nm or 450/50-nm bandpass filter). BV421 is an ideal alternative for V450 as it is approximately ten times brighter with less spillover into the BV510/V500 detector. Please ensure that your instrument’s configurations (lasers and optical filters) are appropriate for this dye.
      altImg
      BV421
      Violet 405 nm
      407 nm
      423 nm
      568418 Rev.1
      Citations & References
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      View product citations for antibody "568418" 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
      568418 Rev. 1

       

      Please refer to Support Documents for Quality Certificates


      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.

      Please refer to Support Documents for Quality Certificates

       

      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.