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PerCP-Cy™5.5 Rat Anti-Mouse NKG2A/C/E
PerCP-Cy™5.5 Rat Anti-Mouse NKG2A/C/E
Two-color flow cytometric analysis of NKG2A/C/E expression on mouse splenocytes. C57BL/6 mouse splenic leucocytes were preincubated with Purified Rat Anti-Mouse CD16/CD32 antibody (Mouse BD Fc Block™) (Cat. No. 553141/553142). The cells were then stained with APC Mouse Anti-Mouse NK-1.1 (Cat. No. 550627/561117) and either PerCP-Cy™5.5 Rat IgG2a, κ Isotype Control (Cat. No. 550765; Left Panel) or PerCP-Cy™5.5 Rat Anti-Mouse NKG2A/C/E antibody (Cat. No. 564384; Right Panel). Two-color flow cytometric contour plots showing the correlated expression of NKG2A/C/E (or Ig Isotype control staining) versus NK-1.1 were derived from gated events with the forward and side light-scatter characteristics of viable splenic leucocytes. Flow cytometric analysis was performed using a BD LSRFortessa™ Cell Analyzer System.
Two-color flow cytometric analysis of NKG2A/C/E expression on mouse splenocytes. C57BL/6 mouse splenic leucocytes were preincubated with Purified Rat Anti-Mouse CD16/CD32 antibody (Mouse BD Fc Block™) (Cat. No. 553141/553142). The cells were then stained with APC Mouse Anti-Mouse NK-1.1 (Cat. No. 550627/561117) and either PerCP-Cy™5.5 Rat IgG2a, κ Isotype Control (Cat. No. 550765; Left Panel) or PerCP-Cy™5.5 Rat Anti-Mouse NKG2A/C/E antibody (Cat. No. 564384; Right Panel). Two-color flow cytometric contour plots showing the correlated expression of NKG2A/C/E (or Ig Isotype control staining) versus NK-1.1 were derived from gated events with the forward and side light-scatter characteristics of viable splenic leucocytes. Flow cytometric analysis was performed using a BD LSRFortessa™ Cell Analyzer System.
Product Details
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BD Pharmingen™
NKG2A/NKG2C/NKG2E; Klrc1/Klrc2/Klrc3; CD159a/CD159c/CD159e; CD159
Mouse (QC Testing)
Rat LEW, also known as Lewis IgG2a, κ
Mouse CD94 and NKG2A Transfected Cell Line
Flow cytometry (Routinely Tested)
0.2 mg/ml
AB_2738784
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 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. Since applications vary, each investigator should titrate the reagent to obtain optimal results.
  2. An isotype control should be used at the same concentration as the antibody of interest.
  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. 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. 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.
  6. 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.
  7. For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
  8. Cy is a trademark of GE Healthcare.
  9. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
564384 Rev. 2
Antibody Details
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20d5

The 20d5 monoclonal antibody specifically recognizes NKG2A, NKG2C, and NKG2E (also known as CD159a, CD159c, and CD159e which are encoded by Klrc1, Klrc2, and Klrc3, respectively) on a subset of NK and NK-T cells in most strains tested (eg, AKR/J, BALB/c, C3H/He, C57BL/6, CBA/J, DBA/1, FVB/N, 129/Sv, NOD, SWR, and most DBA/2 substrains, but not DBA/2J). The NKG2 molecules are a family of lectin-like receptors that form heterodimers with CD94 on the surface of NK cells.  DBA/2J mice do not express CD94, and the lack of CD94 is responsible for the absence of NKG2 expression in this substrain.  NKG2 receptors are also expressed on CD8+ T lymphocytes activated in vivo and in vitro.  The heterodimers of CD94 with NKG2A, C, or E recognize Qa-1, a nonclassical MHC class I antigen, presenting the Qdm peptide.  Studies of CD94/NKG2 heterodimers on human NK cells have demonstrated that the NKG2 components mediate signal transduction for the receptor, with NKG2A being inhibitory and NKG2C being stimulatory. The CD94/NKG2E heterodimer is also thought to be stimulatory.  The mouse NKG2A molecule contains two intracytoplasmic sequences that resemble the ITIM (Immunoreceptor Tyrosine- based Inhibitory Motif) consensus sequence.  NKG2A transcripts have been shown to be up to 20-fold more abundant than NKG2C and NKG2E mRNA in NK cells of adult mice.  The CD94/NKG2 receptors show increased expression on neonatal NK cells compared to the Ly-49 MHC class I receptors, suggesting that CD94/NKG2 receptors and their ligand, Qa-1, may play a role in maintenance of self-tolerance in developing NK cells.  The 20d5 antibody is useful for identification of NK cells expressing functional CD94/NKG2 receptors, in contrast to the non-functional CD94 expressed alone, and it blocks the binding of Qdm-complexed Qa-1b tetramers to CD94/NKG2-transfected CHO cells.  

  

564384 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
564384 Rev.2
Citations & References
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View product citations for antibody "564384" on CiteAb

Development References (7)

  1. Kubota A, Kubota S, Lohwasser S, Mager DL, Takei F. Diversity of NK cell receptor repertoire in adult and neonatal mice. Eur J Immunol. 1999; 163(1):212-216. (Biology). View Reference
  2. Lohwasser S, Hande P, Mager DL, Takei F. Cloning of murine NKG2A, B and C: second family of C-type lectin receptors on murine NK cells. Eur J Immunol. 1999; 29(3):755-761. (Biology). View Reference
  3. McMahon CW, Zajac AJ, Jamieson AM. Viral and bacterial infections induce expression of multiple NK cell receptors in responding CD8(+) T cells. J Immunol. 2002; 169(3):1444-1452. (Clone-specific: Flow cytometry). View Reference
  4. Vance RE, Jamieson AM, Cado D, Raulet DH. Implications of CD94 deficiency and monoallelic NKG2A expression for natural killer cell development and repertoire formation. Proc Natl Acad Sci U S A. 2002; 99(2):868-873. (Clone-specific: Flow cytometry, Fluorescence activated cell sorting). View Reference
  5. Vance RE, Jamieson AM, Raulet DH. Recognition of the class Ib molecule Qa-1(b) by putative activating receptors CD94/NKG2C and CD94/NKG2E on mouse natural killer cells. J Exp Med. 1999; 190(12):1801-1812. (Immunogen: Flow cytometry). View Reference
  6. Vance RE, Kraft JR, Altman JD, Jensen PE, Raulet DH. Mouse CD94/NKG2A is a natural killer cell receptor for the nonclassical major histocompatibility complex (MHC) class I molecule Qa-1(b). J Exp Med. 1998; 188(10):1841-1848. (Biology). View Reference
  7. Yokoyama WM. Natural killer cell receptors. Curr Opin Immunol. 1998; 10(3):298-305. (Biology). View Reference
View All (7) View Less
564384 Rev. 2

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