Description
The 5H10-1 monoclonal antibody specifically recognizes CD8a which is also known as CD8 alpha (CD8α), Ly-2 or Lyt-2. CD8a is an ~34 kDa single pass type I transmembrane glycoprotein that is encoded by Cd8a (CD8 antigen, alpha chain) which belongs to the immunoglobulin superfamily (IgSF). CD8a is comprised of an extracellular region that contains an IgV-like domain followed by a transmembrane region and cytoplasmic tail with a Lck tyrosine kinase binding motif. CD8a is expressed on the cell surface as either a disulfide bond-linked homodimer (CD8αα) or a heterodimer (CD8αβ) when disulfide bonded to CD8b (also known as, CD8 beta/CDβ, Ly-3, or Lyt-3), another type I transmembrane glycoprotein. CD8aβ is expressed on most thymocytes and a subpopulation of mature peripheral TCR αβ T cells including some intraepithelial lymphocytes (IELs). CD8αα is expressed on IELs which include either TCR αβ or TCR γδ T cells as well as on other leucocytes, such as, subsets of NK cells and dendritic cells (DCs). CD8αβ serves as a co-receptor for antigen-stimulated CD8+ T cells by binding to the same peptide:MHC class I complex as the T cell receptor and by helping to trigger the subsequent signaling cascade through Lck activation upon antigen recognition. Through high-affinity binding to nonclassical MHC class-Ib molecules, CD8αα can reportedly function less efficiently as a co-receptor and even repress antigen-stimulated CD8+ T cell 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 Immunofluorescence Applications: The use of a mounting reagent (eg, ProLong® Gold) is highly recommended to maximize the photostability of BV480. For confocal microscopy systems, a 440 nm laser is the optimal excitation source and the recommended emission filter is a 485/20 nm bandpass filter. For epifluorescence microscopes with broad spectrum excitation sources, the recommended excitation and emission filters are 445/20 nm and 485/20 nm bandpass filters, respectively. For specific multicolor imaging applications, the exact filter configurations should be optimized by the end user. For additional instrument/filter configuration information, please visit http://www.bdbiosciences.com/research/cellularimaging.
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. BD Horizon Brilliant Violet 480 is covered by one or more of the following US patents: 8,575,303; 8,354,239.
10. ProLong® is a registered trademark of Thermo Fisher Scientific, Inc. Waltham, MA.
11. BD Horizon Brilliant Violet 480 is covered by one or more of the following US patents: 8,575,303; 8,354,239.
