Description
The 15E2 monoclonal antibody specifically binds to CD369 which is also known as CLEC7A (C-type lectin domain family 7 member A), Dectin-1 (Dendritic cell-associated C-type lectin 1), or BGR (Beta-glucan receptor). CD369 is a type II transmembrane C-type lectin that contains a C-terminal stalk with a carbohydrate recognition domain (CRD) in the extracellular domain, a transmembrane segment, and an ITAM containing cytoplasmic tail. This protein is highly expressed on dendritic cells. It is also expressed on B cells, monocytes, eosinophils, mast cells, macrophages, endothelial cells, and is lowly expressed on some subsets of T cells. CD369 binds to beta-glucan polymers and functions as a pattern recognition receptor in innate immune response to fungal and bacterial pathogens. The 15E2 antibody can reportedly stimulate dendritic cells to costimulate immune responses.
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.
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. When using high concentrations of antibody, background binding of this dye to erythroid fragments produced by ammonium chloride-based lysis, such as with BD Pharm Lyse™ Lysing Buffer (Cat. No. 555899), has been observed when the antibody conjugate was present during the lysis procedure. This may cause nonspecific staining of target cells, such as leukocytes, which have bound the resulting erythroid fragments. This background can be mitigated by any of the following: titrating the antibody conjugate to a lower concentration, fixing samples with formaldehyde, or removing erythrocytes before staining (eg, gradient centrifugation or pre-lysis with wash). This background has not been observed when cells were lysed with BD FACS™ Lysing Solution (Cat. No. 349202) after staining.
2. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
3. 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.
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. An isotype control should be used at the same concentration as the antibody of interest.
7. Since applications vary, each investigator should titrate the reagent to obtain optimal results.
8. Human donor specific background has been observed in relation to the presence of anti-polyethylene glycol (PEG) antibodies, developed as a result of certain vaccines containing PEG, including some COVID-19 vaccines. We recommend use of BD Horizon Brilliant™ Stain Buffer in your experiments to help mitigate potential background. For more information visit https://www.bdbiosciences.com/en-us/support/product-notices.
9. Researchers should determine the optimal concentration of this reagent for their individual applications.
10. 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.
