Preparation And Storage
Recommended Assay Procedures
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).
Note: When using high concentrations of antibody, background binding of this dye to erythroid cell subsets (mature erythrocytes and precursors) has been observed. For researchers studying these cell populations, or in cases where light scatter gating does not adequately exclude these cells from the analysis, this background may be an important factor to consider when selecting reagents for panel(s).
Product Notices
- 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.
- Researchers should determine the optimal concentration of this reagent for their individual applications.
- An isotype control should be used at the same concentration as the antibody of interest.
- 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.
- For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
- Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
- 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.
- Please refer to http://regdocs.bd.com to access safety data sheets (SDS).
- BD Horizon Brilliant Ultraviolet 661 is covered by one or more of the following US patents: 8,110,673; 8,158,444; 8,227,187; 8,575,303; 8,354,239.
Companion Products
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.
Development References (5)
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Joncker NT, Fernandez NC, Treiner E, Vivier E, Raulet DH. NK cell responsiveness is tuned commensurate with the number of inhibitory receptors for self-MHC class I: the rheostat model. J Immunol. 2009; 182(8):4572-4580. (Clone-specific: In vivo exacerbation). View Reference
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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
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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. (Immunogen: Flow cytometry, Fluorescence activated cell sorting). View Reference
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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. (Biology). View Reference
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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
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