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Alexa Fluor™ 647 Mouse Anti-MR1
Alexa Fluor™ 647 Mouse Anti-MR1
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 Alexa Fluor™ 647 Mouse IgG2a, κ Isotype Control (Cat. No. 565357; dashed line histograms) or Alexa Fluor™ 647 Mouse Anti-MR1 antibody (Cat. No. 570026/570107; solid line histograms) at 0.5 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 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 Alexa Fluor™ 647 Mouse IgG2a, κ Isotype Control (Cat. No. 565357; dashed line histograms) or Alexa Fluor™ 647 Mouse Anti-MR1 antibody (Cat. No. 570026/570107; solid line histograms) at 0.5 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.
Product Details
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BD Pharmingen™
HLALS; MHC class I-like antigen MR-1; MHC class-I related-gene protein
Human (QC Testing), Mouse (Tested in Development)
Mouse IgG2a, κ
MR1 Extracellular Domain Complexed with β2m
Flow cytometry (Routinely Tested)
0.2 mg/ml
3140, 15064
Aqueous buffered solution containing protein stabilizer and ≤0.09% sodium azide.
RUO


Preparation And Storage

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. Store undiluted at 4°C and protected from prolonged exposure to light. Do not freeze.

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 cell and BD® CompBeads to ensure that BD® CompBeads are appropriate for your specific cellular application.

Product Notices

  1. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
  2. Alexa Fluor® 647 fluorochrome emission is collected at the same instrument settings as for allophycocyanin (APC).
  3. 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.
  4. Since applications vary, each investigator should titrate the reagent to obtain optimal results.
  5. For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
  6. An isotype control should be used at the same concentration as the antibody of interest.
  7. This product is provided under an intellectual property license between Life Technologies Corporation and BD Businesses. The purchase of this product conveys to the buyer the non-transferable right to use the purchased amount of the product and components of the product in research conducted by the buyer (whether the buyer is an academic or for-profit entity). The buyer cannot sell or otherwise transfer (a) this product (b) its components or (c) materials made using this product or its components to a third party or otherwise use this product or its components or materials made using this product or its components for Commercial Purposes. Commercial Purposes means any activity by a party for consideration and may include, but is not limited to: (1) use of the product or its components in manufacturing; (2) use of the product or its components to provide a service, information, or data; (3) use of the product or its components for therapeutic, diagnostic or prophylactic purposes; or (4) resale of the product or its components, whether or not such product or its components are resold for use in research. For information on purchasing a license to this product for any other use, contact Life Technologies Corporation, Cell Analysis Business Unit Business Development, 29851 Willow Creek Road, Eugene, OR 97402, USA, Tel: (541) 465-8300. Fax: (541) 335-0504.
  8. Species cross-reactivity detected in product development may not have been confirmed on every format and/or application.
  9. Please refer to http://regdocs.bd.com to access safety data sheets (SDS).
  10. Alexa Fluor™ is a trademark of Life Technologies Corporation.
  11. For U.S. patents that may apply, see bd.com/patents.
570107 Rev. 2
Antibody Details
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26.5

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.

570107 Rev. 2
Format Details
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Alexa Fluor™ 647
Alexa Fluor™ 647 Dye is part of the BD red family of dyes. This is a small organic fluorochrome with an excitation maximum (Ex Max) at 653-nm and an emission maximum (Em Max) at 669-nm. Alexa Fluor™ 647 is designed to be excited by the Red laser (627-640 nm) and detected using an optical filter centered near 670-nm (e.g., a 660/20 nm bandpass filter). Please ensure that your instrument’s configurations (lasers and optical filters) are appropriate for this dye.
Alexa Fluor™ 647
Red 627-640 nm
653 nm
669 nm
570107 Rev.2
Citations & References
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View product citations for antibody "570107" 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. (Clone-specific: 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. 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
  6. Lamichhane R, Ussher JE. Expression and trafficking of MR1.. Immunology. 2017; 151(3):270-279. (Biology). View Reference
  7. 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
  8. 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
  9. 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
  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
570107 Rev. 2

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For Research Use Only. Not for use in diagnostic or therapeutic procedures.