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PerCP-Cy™5.5 Mouse Anti-Human CD314 (NKG2D)
PerCP-Cy™5.5 Mouse Anti-Human CD314 (NKG2D)
Flow cytometric analysis of CD314 expression on human peripheral blood lymphocytes. Human whole blood was stained with either PerCP-Cy™5.5 Mouse Anti-Human CD314 antibody (Cat. No. 562364; solid line histogram) or with a PerCP-Cy™5.5 Mouse IgG1, κ Isotype Control (Cat. No. 550795; dashed line histogram). The erythrocytes were lysed with BD Pharm Lyse™ Lysing Buffer (Cat. No. 555899). The fluorescence histograms were derived from events with the forward and side light-scatter characteristics of viable lymphocytes. Flow cytometry was performed using a BD™ LSR II Flow Cytometer System.
Flow cytometric analysis of CD314 expression on human peripheral blood lymphocytes. Human whole blood was stained with either PerCP-Cy™5.5 Mouse Anti-Human CD314 antibody (Cat. No. 562364; solid line histogram) or with a PerCP-Cy™5.5 Mouse IgG1, κ Isotype Control (Cat. No. 550795; dashed line histogram). The erythrocytes were lysed with BD Pharm Lyse™ Lysing Buffer (Cat. No. 555899). The fluorescence histograms were derived from events with the forward and side light-scatter characteristics of viable lymphocytes. Flow cytometry was performed using a BD™ LSR II Flow Cytometer System.
Product Details
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BD Pharmingen™
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
AB_11154225
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 with PerCP-Cy5.5 under optimum conditions, and unconjugated antibody and free PerCP-Cy5.5 were removed. Storage of PerCP-Cy5.5 conjugates in unoptimized diluent is not recommended and may result in loss of signal intensity.

Product Notices

  1. 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).
  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. 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.
  5. 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.
  6. PerCP-Cy5.5–labelled antibodies can be used with FITC- and R-PE–labelled reagents in single-laser flow cytometers with no significant spectral overlap of PerCP-Cy5.5, FITC, and R-PE fluorescence.
  7. For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
  8. PerCP-Cy5.5 is optimized for use with a single argon ion laser emitting 488-nm light. Because of the broad absorption spectrum of the tandem fluorochrome, extra care must be taken when using dual-laser cytometers, which may directly excite both PerCP and Cy5.5™. We recommend the use of cross-beam compensation during data acquisition or software compensation during data analysis.
  9. Cy is a trademark of GE Healthcare.
  10. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
562364 Rev. 2
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.

562364 Rev. 2
Format Details
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PerCP-Cy5.5
PerCP-Cy5.5 dye is part of the BD blue family of dyes. This tandem fluorochrome is comprised of a fluorescent protein complex (PerCP) with an excitation maximum (Ex Max) of 482 nm and an acceptor dye with an emission maximum (Em Max) at 676 nm. PerCP-Cy5 is designed to be excited by the blue laser (488-nm) and detected using an optical filter centered near 680 nm (e.g., a 695/40 nm bandpass filter). The donor dye can be partially excited by the Violet (405-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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PerCP-Cy5.5
Blue 488 nm
482 nm
676 nm
562364 Rev.2
Citations & References
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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, 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: (Co)-stimulation, Flow cytometry, Functional assay). 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
View All (5) View Less
562364 Rev. 2

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