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Biotin Rat Anti-Mouse CD94
Biotin Rat Anti-Mouse CD94

Expression of CD94 on mouse splenic NK cells. C57BL/6 splenocytes were preincubated with Mouse BD Fc Block™ purified anti-mouse CD16/CD32 mAb 2.4G2 (Cat. no. 553141/553142), then simultaneously stained with biotinconjugated rat IgG2a isotype control mAb R35-95 (Cat. no. 553928, Left panel) or biotinylated mAb 18d3 (Right panel) and APC-conjugated anti- mouse NK-1.1 mAb PK136 (Cat. no. 550627), followed by Streptavidin-PE (Cat. no. 554061). The CD94[dim] and CD94[bright] subpopulations of NK cells are observed in all strains tested, although the relative proportions of the two subsets may vary. Flow cytometry was performed on a BD FACSCalibur™ flow cytometry system.

Expression of CD94 on mouse splenic NK cells. C57BL/6 splenocytes were preincubated with Mouse BD Fc Block™ purified anti-mouse CD16/CD32 mAb 2.4G2 (Cat. no. 553141/553142), then simultaneously stained with biotinconjugated rat IgG2a isotype control mAb R35-95 (Cat. no. 553928, Left panel) or biotinylated mAb 18d3 (Right panel) and APC-conjugated anti- mouse NK-1.1 mAb PK136 (Cat. no. 550627), followed by Streptavidin-PE (Cat. no. 554061). The CD94[dim] and CD94[bright] subpopulations of NK cells are observed in all strains tested, although the relative proportions of the two subsets may vary. Flow cytometry was performed on a BD FACSCalibur™ flow cytometry system.

Product Details
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BD Pharmingen™
Mouse (QC Testing)
Rat LEW, also known as Lewis IgG2a, κ
Transfected cell line
Flow cytometry (Routinely Tested), Immunohistochemistry-frozen (Not Recommended)
0.5 mg/ml
AB_393878
Aqueous buffered solution containing ≤0.09% sodium azide.
RUO


Preparation And Storage

The monoclonal antibody was purified from tissue culture supernatant or ascites by affinity chromatography. The antibody was conjugated with biotin under optimum conditions, and unreacted biotin was removed. Store undiluted at 4°C and protected from prolonged exposure to light. Do not freeze.

Recommended Assay Procedures

We have found that the use of Mouse BD Fc Block, purified anti-mouse CD16/CD32 mAb 2.4G2 (Cat. no. 553141/553142) reduces the non-specific staining of non-NK cells by this biotin conjugate. This antibody is not useful for immunohistochemical staining of frozen sections.

Product Notices

  1. Since applications vary, each investigator should titrate the reagent to obtain optimal results.
  2. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
  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.
550773 Rev. 5
Antibody Details
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18d3

The 18d3 monoclonal antibody specifically recognizes CD94 on all NK cells, NK1.1- or DX5-positive T lymphocytes (NK-T cells), and a subset of CD8-positive T lymphocytes in most strains tested (eg, A/J, 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). DBA/2J mice do not express CD94.3 CD94 is also expressed on CD8+ T lymphocytes activated in vivo. CD94 is a type-II transmembrane protein with an extracellular lectin-like domain and a short cytoplasmic tail which is not believed to have any signalling function.  Heterodimers of CD94 with NKG2A, NKG2C, or NKG2E recognize Qa-1 (a non-classical MHC class I antigen) presenting the Qdm peptide.  Studies on CD94/NKG2 heterodimers on human NK cells have demonstrated that the NKG2 components mediate signal  transduction for the receptor, NKG2A being inhibitory and NKG2C being  stimulatory.  Similarly, the mouse NKG2A molecule contains two intracytoplasmic sequences which resemble the ITIM (Immunoreceptor Tyrosine-based Inhibitory Motif) consensus sequence. CD94/NKG2 receptors appear on fetal NK cells before 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.

550773 Rev. 5
Format Details
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Biotin
Biotin is a ubiquitous co-factor (also known as Vitamin B7) that has many properties that make it extremely useful for molecular biology. Biotin has an extremely high affinity for the Avidin family of proteins (Kd = 10-15 M), making it the perfect tool to link two molecules. Biotin labeled antibodies can be combined with any number of Avidin-conjugated probes in order to customize an assay to a particular need. This is especially useful in the case of magnetic cell separation using streptavidin/magnetic bead conjugates, or in the case of flow cytometry using streptavidin/fluorophore conjugates.
Biotin
550773 Rev.5
Citations & References
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Development References (8)

  1. Ho EL, Heusel JW, Brown MG. Murine Nkg2d and Cd94 are clustered within the natural killer complex and are expressed independently in natural killer cells. Proc Natl Acad Sci U S A. 1998; 95(11):6320-6325. (Biology). View Reference
  2. 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. (Biology). View Reference
  3. Sivakumar PV, Gunturi A, Salcedo M, et al. Cutting edge: expression of functional CD94/NKG2A inhibitory receptors on fetal NK1.1+Ly-49- cells: a possible mechanism of tolerance during NK cell development. J Immunol. 1999; 162(12):6976-6980. (Biology). View Reference
  4. Toomey JA, Salcedo M, Cotterill LA. Stochastic acquisition of Qa1 receptors during the development of fetal NK cells in vitro accounts in part but not in whole for the ability of these cells to distinguish between class I-sufficient and class I-deficient targets. J Immunol. 1999; 163(6):3176-3184. (Biology). View Reference
  5. 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. (Biology). View Reference
  6. 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). View Reference
  7. 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
  8. Yokoyama WM. Natural killer cell receptors . Curr Opin Immunol. 1998; 10:298-305. (Biology).
View All (8) View Less
550773 Rev. 5

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