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PE-CF594 Mouse Anti-Human HLA-DR
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PE-CF594 Mouse Anti-Human HLA-DR
Multiparameter flow cytometric analysis of HLA-DR expression on Human peripheral blood leucocyte populations. Human whole blood was stained with either BD Horizon™ PE-CF594 Mouse IgG2a, κ Isotype Control (Cat. No. 562306; Left Plot) or BD Horizon™ PE-CF594 Mouse Anti-Human HLA-DR antibody (Cat. No. 568651/568652; Right Plot). Erythrocytes were lysed with BD FACS Lysing™ Solution (Cat. No. 349202). The bivariate pseudocolor density plot showing the correlated expression of HLA-DR (or Ig Isotype control staining) versus side-light scatter (SSC-A) signals was derived from gated events with the side and forward light-scatter characteristics of intact leucocyte populations. Flow cytometry and data analysis were performed using a BD LSRFortessa™ X-20 Cell Analyzer System and FlowJo™ software.
Multiparameter flow cytometric analysis of HLA-DR expression on Human peripheral blood leucocyte populations. Human whole blood was stained with either BD Horizon™ PE-CF594 Mouse IgG2a, κ Isotype Control (Cat. No. 562306; Left Plot) or BD Horizon™ PE-CF594 Mouse Anti-Human HLA-DR antibody (Cat. No. 568651/568652; Right Plot). Erythrocytes were lysed with BD FACS Lysing™ Solution (Cat. No. 349202). The bivariate pseudocolor density plot showing the correlated expression of HLA-DR (or Ig Isotype control staining) versus side-light scatter (SSC-A) signals was derived from gated events with the side and forward light-scatter characteristics of intact leucocyte populations. Flow cytometry and data analysis were performed using a BD LSRFortessa™ X-20 Cell Analyzer System and FlowJo™ software.
Product Details
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BD Horizon™
MHC class II antigen; HLA class II histocompatibility antigen
Human (QC Testing), Rhesus, Cynomolgus, Baboon (Tested in Development)
Mouse IgG2a, κ
Human lymphoblastoid B-cell line RPMI 8866
Flow cytometry (Routinely Tested)
5 µl
Aqueous buffered solution containing BSA and ≤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.

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.

Product Notices

  1. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
  2. Source of all serum proteins is from USDA inspected abattoirs located in the United States.
  3. An isotype control should be used at the same concentration as the antibody of interest.
  4. Species cross-reactivity detected in product development may not have been confirmed on every format and/or application.
  5. Please observe the following precautions: Absorption of visible light can significantly alter the energy transfer occurring in any tandem fluorochrome conjugate; therefore, we recommend that special precautions be taken (such as wrapping vials, tubes, or racks in aluminum foil) to prevent exposure of conjugated reagents, including cells stained with those reagents, to room illumination.
  6. 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.
  7. This reagent has been pre-diluted for use at the recommended Volume per Test. We typically use 1 × 10^6 cells in a 100-µl experimental sample (a test).
  8. For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
  9. When excited by the yellow-green (561-nm) laser, the fluorescence may be brighter than when excited by the blue (488-nm) laser.
  10. This product is provided under an Agreement between BIOTIUM and BD Biosciences. The manufacture, use, sale, offer for sale, or import of this product is subject to one or more patents or pending applications owned or licensed by Biotium, Inc. 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.
  11. Because of the broad absorption spectrum of the tandem fluorochrome, extra care must be taken when using multi-laser cytometers, which may directly excite both PE and CF™594.
  12. Please refer to http://regdocs.bd.com to access safety data sheets (SDS).
  13. Texas Red is a registered trademark of Molecular Probes, Inc., Eugene, OR.
  14. CF™ is a trademark of Biotium, Inc.
568651 Rev. 1
Antibody Details
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L243

The L243 monoclonal antibody specifically binds to HLA-DR, a major histocompatibility complex (MHC) class II antigen. HLA-DR antigens are encoded by genes within the Human Leukocyte Antigen (HLA) Complex located on chromosome 6. HLA-DR is a transmembrane heterodimeric glycoprotein composed of an α chain (36 kDa) and a β subunit (27 kDa) expressed primarily on antigen presenting cells including B cells, dendritic cells, monocytes, macrophages, Langerhans cells, and thymic epithelial cells. HLA-DR is also expressed on activated T cells. This molecule plays a major role in mediating cellular interactions during antigen presentation to CD4-positive T cells.

568651 Rev. 1
Format Details
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PE-CF594
BD Horizon™ PE-CF594 dye is a part of the BD PE family of dyes. This tandem fluorochrome is comprised of a R-Phycoerythrin (PE) donor that has excitation maxima (Ex Max) of 496-nm and 566-nm and an acceptor dye with an emission maximum (Em Max) at 615-nm. PE-CF594, driven by BD innovation, is designed to be excited by the blue (488-nm), Green (532-nm) and yellow-green (561-nm) lasers and detected using an optical filter centered near 615 nm (e.g., a 610/20-nm bandpass filter). The donor dye can be excited by the Blue (488-nm), Green (532-nm) and yellow-green (561-nm) lasers and the acceptor dye can be excited by the green (532-nm) laser resulting in cross-laser excitation and fluorescence spillover. Please ensure that your instrument’s configurations (lasers and optical filters) are appropriate for this dye.
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PE-CF594
Yellow-Green 488 nm, 532 nm, 561 nm
496 nm, 566 nm
615 nm
568651 Rev.1
Citations & References
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View product citations for antibody "568651" on CiteAb

Development References (12)

  1. Autissier P, Soulas C, Burdo TH, Williams KC. Evaluation of a 12-color flow cytometry panel to study lymphocyte, monocyte, and dendritic cell subsets in humans.. Cytometry A. 2010; 77(5):410-9. (Clone-specific: Flow cytometry). View Reference
  2. Brodsky FM. A matrix approach to human class II histocompatibility antigens: reactions of four monoclonal antibodies with the products of nine haplotypes.. Immunogenetics. 1984; 19(3):179-94. (Clone-specific: Blocking, Immunoprecipitation, Radioimmunoassay). View Reference
  3. Charles N, Hardwick D, Daugas E, Illei GG, Rivera J. Basophils and the T helper 2 environment can promote the development of lupus nephritis.. Nat Med. 2010; 16(6):701-7. (Clone-specific: Flow cytometry). View Reference
  4. Engleman EG, Warnke R, Fox RI, Dilley J, Benike CJ, Levy R. Studies of a human T lymphocyte antigen recognized by a monoclonal antibody.. Proc Natl Acad Sci USA. 1981; 78(3):1791-5. (Clone-specific: Immunohistochemistry). View Reference
  5. Fujita H, Nograles KE, Kikuchi T, Gonzalez J, Carucci JA, Krueger JG. Human Langerhans cells induce distinct IL-22-producing CD4+ T cells lacking IL-17 production.. Proc Natl Acad Sci U S A. 2009; 106(51):21795-800. (Clone-specific: Flow cytometry). View Reference
  6. Harper D, Gollackner B, Xu Y, et al. In vitro and in vivo investigation of a novel monoclonal antibody to plasma cells (W5 mAb).. Xenotransplantation. 2004; 11(1):78-90. (Clone-specific: Immunoprecipitation). View Reference
  7. Lampson LA, Levy R. Two populations of Ia-like molecules on a human B cell line.. J Immunol. 1980; 125(1):293-9. (Immunogen: Blocking, Immunoprecipitation, Radioimmunoassay). View Reference
  8. Ren Z, Wang J, Zhu W, et al. Spontaneous transformation of adult mesenchymal stem cells from cynomolgus macaques in vitro.. Exp Cell Res. 2011; 317(20):2950-7. (Clone-specific: Flow cytometry). View Reference
  9. Stumptner-Cuvelette P, Morchoisne S, Dugast M, et al. HIV-1 Nef impairs MHC class II antigen presentation and surface expression.. Proc Natl Acad Sci U S A. 2001; 98(21):12144-9. (Clone-specific: Flow cytometry). View Reference
  10. Tomkinson BE, Wagner DK, Nelson DL, Sullivan JL. Activated lymphocytes during acute Epstein-Barr virus infection.. J Immunol. 1987; 139(11):3802-7. (Clone-specific: Flow cytometry). View Reference
  11. Wang RF, Wang X, Atwood AC, Topalian SL, Rosenberg SA. Cloning genes encoding MHC class II-restricted antigens: mutated CDC27 as a tumor antigen.. Science. 1999; 284(5418):1351-4. (Clone-specific: Blocking). View Reference
  12. Weisgrau KL, Vosler LJ, Pomplun NL, et al. Neutrophil progenitor populations of rhesus macaques.. J Leukoc Biol. 2019; 105(1):113-121. (Clone-specific: Cell separation, Flow cytometry). View Reference
View All (12) View Less
568651 Rev. 1

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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.