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BV605 Rat Anti-Mouse NKG2A/C/E
BV605 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 BD Horizon™ BV605 Rat IgG2a, κ Isotype Control (Cat. No. 563144; Left Panel) or BD Horizon BV605 Rat Anti-Mouse NKG2A/C/E antibody (Cat. No. 564382; 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™ LSR II Flow Cytometer 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 BD Horizon™ BV605 Rat IgG2a, κ Isotype Control (Cat. No. 563144; Left Panel) or BD Horizon BV605 Rat Anti-Mouse NKG2A/C/E antibody (Cat. No. 564382; 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™ LSR II Flow Cytometer System.
Product Details
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BD Horizon™
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_2738782
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 BD Horizon™ BV605 under optimum conditions, and unconjugated antibody and free BD Horizon™ BV605 were removed.

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. Although every effort is made to minimize the lot-to-lot variation in the efficiency of the fluorochrome energy transfer, differences in the residual emission from BD Horizon™ BV421 may be observed. Therefore, we recommend that individual compensation controls be performed for every BD Horizon™ BV605 conjugate.
  6. CF™ is a trademark of Biotium, Inc.
  7. For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
  8. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
564382 Rev. 3
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.  

This antibody is conjugated to BD Horizon BV605 which is part of the BD Horizon Brilliant™ Violet family of dyes. With an Ex Max of 407-nm and Em Max of 602-nm, BD Horizon BV605 can be excited by a violet laser and detected with a standard 610/20-nm filter set. BD Horizon BV605 is a tandem fluorochrome of BD Horizon BV421 and an acceptor dye with an Em max at 605-nm. Due to the excitation of the acceptor dye by the green (532 nm) and yellow-green (561 nm) lasers, there will be significant spillover into the PE and BD Horizon PE-CF594 detectors off the green or yellow-green lasers. BD Horizon BV605 conjugates are very bright, often exhibiting brightness equivalent to PE conjugates and can be used as a third color off of the violet laser.

For optimal and reproducible results, BD Horizon Brilliant Stain Buffer should be used anytime two or more BD Horizon Brilliant dyes are used in the same experiment.  Fluorescent dye interactions may cause staining artifacts which may affect data interpretation.  The BD Horizon Brilliant Stain Buffer was designed to minimize these interactions.  More information can be found in the Technical Data Sheet of the BD Horizon Brilliant Stain Buffer (Cat. No. 563794).

564382 Rev. 3
Format Details
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BV605
The BD Horizon Brilliant Violet™ 605 (BV605) dye is part of the BD Horizon Brilliant Violet™ family of dyes. This tandem fluorochrome is comprised of a BV421 donor with an excitation maximum (Ex Max) of 407-nm and an acceptor dye with an emission maximum (Em Max) at 605-nm. BV605, driven by BD innovation, is designed to be excited by the violet laser (405-nm) and detected using an optical filter centered near 610-nm (e.g., a 610/20-nm bandpass filter). The acceptor dye can be excited by the yellow-green (561-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.
altImg
BV605
Violet 405 nm
407 nm
605 nm
564382 Rev.3
Citations & References
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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: ELISA, 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
564382 Rev. 3

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