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RY586 Mouse Anti-Human CD314 (NKG2D)
RY586 Mouse Anti-Human CD314 (NKG2D)
Two-color flow cytometric analysis of CD314 (NKG2D) expression on Human peripheral blood lymphocytes. Human whole blood was stained with APC Mouse Anti-Human CD56 (NCAM-1) [Cat. No. 555518] and with either BD Horizon™ RY586 Mouse IgG1, κ Isotype Control (Cat. No. 568097; Left Plot) or BD Horizon™ RY586 Mouse Anti-Human CD314 (NKG2D) antibody (Cat. No. 568499/568500; Right Plot). The erythrocytes were lysed with BD FACS™ Lysing Solution (Cat. No. 349202). The bivariate pseudocolor density plot showing the correlated expression of CD314 (NKG2D) [or Ig Isotype control staining] versus CD56 (NCAM-1) was derived from gated events with the forward and side light-scatter characteristics of intact lymphocytes. Flow cytometry and data analysis were performed using a BD LSRFortessa™ X-20 Cell Analyzer System and FlowJo™ software.
RY586 Mouse Anti-Human CD314 (NKG2D)
Multiparameter flow cytometric analysis of CD314 (NKG2D) expression on Human peripheral blood leucocytes. Human whole blood was stained with either BD Horizon™ RY586 Mouse IgG1, κ Isotype Control (Cat. No. 568097; Left Plot) or BD Horizon™ RY586 Mouse Anti-Human CD314 (NKG2D) antibody (Cat. No. 568499/568500; Right Plot). The erythrocytes were lysed with BD FACS™ Lysing Solution (Cat. No. 349202). The bivariate pseudocolor density plot showing the correlated expression of CD314 (NKG2D) [or Ig Isotype control staining] versus side light-scatter (SSC-A) signals was derived from gated events with the forward and side light-scatter characteristics of intact leucocytes. Flow cytometry and data analysis were performed using a BD LSRFortessa™ X-20 Cell Analyzer System and FlowJo™ software.
Two-color flow cytometric analysis of CD314 (NKG2D) expression on Human peripheral blood lymphocytes. Human whole blood was stained with APC Mouse Anti-Human CD56 (NCAM-1) [Cat. No. 555518] and with either BD Horizon™ RY586 Mouse IgG1, κ Isotype Control (Cat. No. 568097; Left Plot) or BD Horizon™ RY586 Mouse Anti-Human CD314 (NKG2D) antibody (Cat. No. 568499/568500; Right Plot). The erythrocytes were lysed with BD FACS™ Lysing Solution (Cat. No. 349202). The bivariate pseudocolor density plot showing the correlated expression of CD314 (NKG2D) [or Ig Isotype control staining] versus CD56 (NCAM-1) was derived from gated events with the forward and side light-scatter characteristics of intact lymphocytes. Flow cytometry and data analysis were performed using a BD LSRFortessa™ X-20 Cell Analyzer System and FlowJo™ software.
Multiparameter flow cytometric analysis of CD314 (NKG2D) expression on Human peripheral blood leucocytes. Human whole blood was stained with either BD Horizon™ RY586 Mouse IgG1, κ Isotype Control (Cat. No. 568097; Left Plot) or BD Horizon™ RY586 Mouse Anti-Human CD314 (NKG2D) antibody (Cat. No. 568499/568500; Right Plot). The erythrocytes were lysed with BD FACS™ Lysing Solution (Cat. No. 349202). The bivariate pseudocolor density plot showing the correlated expression of CD314 (NKG2D) [or Ig Isotype control staining] versus side light-scatter (SSC-A) signals was derived from gated events with the forward and side light-scatter characteristics of intact leucocytes. 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™
CD314; KLRK1; KLR; NKG2D; NKG2-D; NK cell receptor D
Human (QC Testing)
Mouse RBF/ DnJ IgG1, κ
NKL cells
Flow cytometry (Routinely Tested)
5 µl
VIII 80432
22914
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

  1. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
  2. 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).
  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. 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. Please refer to http://regdocs.bd.com to access safety data sheets (SDS).
  8. CF™ is a trademark of Biotium, Inc.
568499 Rev. 1
Antibody Details
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1D11

The 1D11 monoclonal antibody specifically binds to NKG2D, a 42 kDa type II transmembrane glycoprotein that is also known as CD314 and KLRK1. NKG2D is a member of the C-type lectin family and is expressed on human NK cells. This activating receptor binds strongly to several ligands including MICA and MICB and ULBP-1, -2, and -3 proteins that are expressed by different target cell types. Different from natural cytotoxicity receptor (NCR), NKG2D expression is not confined to NK cells. It is also expressed on virtually all TCR γ/δ+ and CD8+TCR α/β+ T cells. NKG2D functions as a triggering receptor involved in natural cytotoxicity mediated by normal NK cells against a variety of tumors or normal target cells. Importantly, NKG2D can complement the role of NCR in tumor cell lysis. Remarkably, the combined maskings of NCR and NKG2D can reportedly lead to a complete inhibition of NK-mediated lysis of all tumor or normal cells. The 1D11 antibody can reportedly block or stimulate the function of NKG2D-positive cells.

568499 Rev. 1
Format Details
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RY586
The BD Horizon RealYellow™ 586 (RY586) Dye is part of the BD family of yellow-green dyes. It is a small organic fluorochrome with an excitation maximum (Ex Max) at 565-nm and an emission maximum (Em Max) at 586-nm. Driven by BD innovation, RY586 can be used on both spectral and conventional cytometers and is designed to be excited by the Yellow-Green laser (561-nm) with minimal excitation by the 488-nm Blue laser. For conventional instruments equipped with a Yellow-Green laser (561-nm), RY586 can be used as an alternative to PE and we recommend using an optical filter centered near 586-nm (eg, a 586/15-nm bandpass filter). For spectral instruments equipped with a Yellow-Green laser (561-nm), it can be used in conjunction with PE. Compared to PE, RY586 is similar in brightness, minimal spillover into Blue detectors, and increased spillover into the 610/20-nm (PE-CF594) detector. Please ensure that your instrument configuration (lasers and optical filters) is appropriate for this dye.
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RY586
Yellow-Green 561 nm
564 nm
586 nm
568499 Rev.1
Citations & References
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View product citations for antibody "568499" on CiteAb

Development References (5)

  1. Bauer S, Groh V, Wu J, et al. Activation of NK cells and T cells by NKG2D, a receptor for stress-inducible MICA. Science. 1999; 285(5428):727-729. (Immunogen: Blocking, Flow cytometry, Functional assay, Immunoprecipitation, Inhibition). View Reference
  2. Groh V, Bruhl A, El-Gabalawy H, Nelson JL, Spies T. Stimulation of T cell autoreactivity by anomalous expression of NKG2D and its MIC ligands in rheumatoid arthritis. Proc Natl Acad Sci U S A. 2003; 100(16):9452-9457. (Clone-specific: Flow cytometry, Immunohistochemistry). View Reference
  3. Groh V, Rhinehart R, Randolph-Habecker J, Topp MS, Riddell SR, Spies T. Costimulation of CD8alphabeta T cells by NKG2D via engagement by MIC induced on virus-infected cells. Nat Immunol. 2001; 2(3):255-260. (Clone-specific: Blocking, (Co)-stimulation, Immunohistochemistry, Inhibition). View Reference
  4. Roberts AI, Lee L, Schwarz E, et al. NKG2D receptors induced by IL-15 costimulate CD28-negative effector CTL in the tissue microenvironment. J Immunol. 2001; 167(10):5527-5530. (Clone-specific: Flow cytometry, Stimulation). View Reference
  5. Steinle A, Li P, Morris DL, et al. Interactions of human NKG2D with its ligands MICA, MICB, and homologs of the mouse RAE-1 protein family. Immunogenetics. 2001; 53(4):279-287. (Clone-specific: Blocking). View Reference
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568499 Rev. 1

 

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