Description
The NFLD.D1 monoclonal antibody is specific for the HLA-DR4 serogroup. This panspecific HLA-DR4 antibody binds to an epitope expressed in the extracellular beta-2 (β2) domain of all HLA-DRB1 beta chain variants encoded by HLA-DRB1*04 alleles. The epitope recognized by the NFLD.D1 antibody depends on both a leucine present at sequence position 180 and a threonine at position 181. HLA-DR antigens are heterodimers comprised of type I transmembrane glycoprotein alpha and beta subunits that contain two extracellular domains, transmembrane regions, and cytoplasmic tails. The ~34 kDa HLA-DR alpha (HLA-DRα) chain is encoded by HLA-DRA whereas the alternative ~28 kDa HLA-DR beta (HLA-DRβ) chains are encoded by one of the four different HLA-DRB loci (HLA-DRB1,3,4, and 5) that are located within the Human Leukocyte Antigen (HLA) Complex of chromosome 6. HLA-DR is variably expressed on antigen-presenting cells (APCs), such as dendritic cells, B lymphocytes, monocytes, macrophages, and thymic epithelial cells as well as by activated T cells, epithelial cells, inflammatory fibroblasts, and tumor cells. Polymorphisms in the HLA-DRβ chains allow for variability in peptide binding and the presentation of self and foreign peptide antigens to CD4+ T lymphocytes that generate and regulate adaptive immune responses. The NFLD.D1 antibody can be especially useful along with other HLA-DRβ allotype-specific antibodies in applications such as immunofluorescence and flow cytometry for analyzing allelic HLA-DRB expression patterns by individual cells within heterogeneous cell populations. Some HLA-DRB1*04 alleles or their aberrant expression patterns have been associated with susceptibility to diseases including rheumatoid arthritis.
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).
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. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
2. Please refer to http://regdocs.bd.com to access safety data sheets (SDS).
3. For U.S. patents that may apply, see bd.com/patents.
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. Since applications vary, each investigator should titrate the reagent to obtain optimal results.
6. 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.
7. For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
8. An isotype control should be used at the same concentration as the antibody of interest.
