The 16a11 monoclonal antibody specifically recognizes the Mouse CD159a alloantigen that is also known as NKG2A alloantigen (NKG2A[B6] or NKG2AB6) which is expressed on subsets of C57BL/6 mouse NK, NK-T, or activated CD8+ T cells. CD159a (NKG2AB6) is an ~40 kDa single-pass type II transmembrane glycoprotein that is encoded by Klrc1 (Killer cell lectin-like receptor subfamily C member 1). This NKG2 receptor is comprised of an extracellular region with one C-type lectin domain followed by a transmembrane segment and a cytoplasmic tail with two immunoreceptor tyrosine-based inhibitory motifs (ITIMs). It is expressed on the cell surface as a heterodimer that is disulfide bonded to CD94, an invariant type II C-type lectin-like transmembrane glycoprotein. The 16a11 antibody does not recognize NKG2A[BALB], the NKG2A alloantigen expressed by BALB/c mouse leucocytes. This antibody neither crossreacts with other NKG2 family members, NKG2C (NKG2C[B6]) nor NKG2E (NKG2E[B6]) alloantigens, nor CD94. Heterodimeric complexes of CD94 with either NKG2A, C, or E recognize Qa-1, a non-classical self-MHC class I antigen, presenting the Qdm signal peptide. This recognition endows NK cells with the capacity to detect cells with abnormal MHC class I expression that may result from cellular transformation or viral infection. Ligand-bound CD159a (NKG2AB6):CD94 can play a role in the inhibition of NK cell-mediated target cell lysis. When costaining cells with the 20d5 monoclonal antibody that is specific for mouse NKG2A/C/E receptors, crossblocking was reportedly minimized by first incubating cells with the 16a11 antibody and followed by the 20d5 antibody.
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. For optimal and reproducible results, BD Horizon Brilliant™ Stain Buffer should be used anytime BD Horizon Brilliant dyes are used in a multicolor flow cytometry panel. Fluorescent dye interactions may cause staining artifacts which may affect data interpretation. The BD Horizon Brilliant Stain Buffer was designed to minimize these interactions. When BD Horizon Brilliant Stain Buffer is used in in the multicolor panel, it should also be used in the corresponding compensation controls for all dyes to achieve the most accurate compensation. For the most accurate compensation, compensation controls created with either cells or beads should be exposed to BD Horizon Brilliant Stain Buffer for the same length of time as the corresponding multicolor panel. More information can be found in the Technical Data Sheet of the BD Horizon Brilliant Stain Buffer (Cat. No. 563794/566349) or the BD Horizon Brilliant Stain Buffer Plus (Cat. No. 566385).
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 that minimize unconjugated dye and antibody.
1. The production process underwent stringent testing and validation to assure that it generates a high-quality conjugate with consistent performance and specific binding activity. However, verification testing has not been performed on all conjugate lots.
2. Researchers should determine the optimal concentration of this reagent for their individual applications.
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. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
7. BD Horizon Brilliant Stain Buffer is covered by one or more of the following US patents: 8,110,673; 8,158,444; 8,575,303; 8,354,239.
8. Please refer to http://regdocs.bd.com to access safety data sheets (SDS).
9. 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.
10. CF™ is a trademark of Biotium, Inc.
11. 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.
12. BD Horizon Brilliant Violet 605 is covered by one or more of the following US patents: 8,110,673; 8,158,444; 8,227,187; 8,455,613; 8,575,303; 8,354,239.
