Description
The CH-L monoclonal antibody specifically binds to CD158b proteins. These proteins are 50-58 kDa type I glycoproteins that belong to the Killer cell immunoglobulin-like receptor (KIR) family: (KIR2DL2/L3/S2). They are also known as CD158b1 (KIR2DL2; NKAT-6; p58.2), CD158b2 (KIR2DL3; NKAT-2; p58.2), or CD158j (KIR2DS2; NKAT-5; p50.2). The CD158b molecules are composed of two extracellular Ig-like domains, and a transmembrane region. CD158b1 and CD158b2 also possess long (84 or 76 amino acids, respectively) cytoplasmic tails with two immunoreceptor tyrosine-based inhibition motifs (ITIM) whereas CD158j has a short (39 amino acid) cytoplasmic tail that lacks the ITIM motif. CD158b molecules are expressed on NK cells and subsets of TCR αβ+ cells or TCR γδ+ cells. Ligand- or CH-L antibody-bound CD158b1 or CD158b2 can reportedly inhibit cytolytic NK and T cell responses to various stimuli including certain target cells expressing MHC class I ligands encoded by HLA-C alleles (Cw 1, 3, 7 and 8). CD158j reportedly can enhance some cellular cytolytic responses.
BD™ CompBeads can be used as surrogates to assess fluorescence spillover (Compensation). When fluorochrome conjugated antibodies are bound to 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 CompBead to ensure that BD Comp beads are appropriate for your specific cellular application. 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. 563794/566349) or the BD Horizon Brilliant Stain Buffer Plus (Cat. No. 566385). For optimal results, it is recommended to perform 2 washes after staining with antibodies. Cells may be prepared, stained with antibodies and washed twice with wash buffer per established protocols for immunofluorescence staining, prior to acquisition on a flow cytometer. Performing fewer than the recommended wash steps may lead to increased spread of the negative population.
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™ BB515 under optimum conditions and unconjugated antibody was removed.
1. This reagent has been pre-diluted for use at the recommended Volume per Test. We typically use 1 × 10^6 cells in a 100-µl experimental sample (a test).
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. For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
5. 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.
6. Please refer to http://regdocs.bd.com to access safety data sheets (SDS).
7. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
