Multiparameter flow cytometric analysis of HLA-DR expression on Human peripheral blood leucocyte populations. Human whole blood was stained with either BD Horizon™ R718 Mouse IgG2a, κ Isotype Control (Cat. No. 566949; Left Plot) or BD Horizon™ R718 Mouse Anti-Human HLA-DR antibody (Cat. No. 568578/568579; 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.
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Multiparameter flow cytometric analysis of HLA-DR expression on Human peripheral blood leucocyte populations. Human whole blood was stained with either BD Horizon™ R718 Mouse IgG2a, κ Isotype Control (Cat. No. 566949; Left Plot) or BD Horizon™ R718 Mouse Anti-Human HLA-DR antibody (Cat. No. 568578/568579; 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
BD Horizon™
MHC class II antigen; HLA class II histocompatibility antigen
Human (QC Testing)
Mouse IgG2a, κ
Human lymphoblastoid B-cell line RPMI 8866
Flow cytometry (Routinely Tested)
5 µl
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.
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
- Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
- 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).
- 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.
- Please refer to http://regdocs.bd.com to access safety data sheets (SDS).
- 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.
- 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.
- Alexa Fluor™ is a trademark of Life Technologies Corporation.
Companion Products
Stain Buffer (FBS) RUO
Size
500 mL
Cat No.
554656
Stain Buffer (BSA) RUO
Size
500 mL
Cat No.
554657
R718 Mouse IgG2a, κ Isotype Control RUO
Size
50 µg
Cat No.
566949
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Lysing Solution 10X Concentrate IVD
Size
100 mL
Cat No.
349202
Lysing Buffer RUO
Size
100 mL
Cat No.
555899
Antibody Details
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.
Format Details
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.
R718
Red 627-640 nm
695 nm
718 nm
Citations & References
View product citations for antibody "568579" on CiteAb
Development References (9)
-
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
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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
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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
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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
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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
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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
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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
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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
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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
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.