The M1/42 monoclonal antibody (mAb) specifically recognizes intact Histocompatibility-2 Region (H-2) class I antigens (Ags), also known as mouse Major Histocompatibility Complex class I (MHC-I) Ags. This mAb reportedly binds to one or more of the H-2 Ags (H-2K, H-2D, and likely H-2L) that are primarily expressed on most nucleated somatic cells of hematopoietic and nonhematopoietic origin. H-2 class I molecules are comprised of a ~45-50 kDa type I transmembrane heavy chain glycoprotein that is noncovalently linked to ~13 kDa β2-microglobulin (β2-m). The extracellular region of the polymorphic heavy chains is comprised of three globular domains (α1, α2, and α3), followed by a transmembrane region and a cytoplasmic tail. Sequence variations are manifest in regions of the α1 and α2 domains that line the peptide-binding cleft involved in Ag presentation. The M1/42 mAb reportedly binds to H-2 class I Ags from multiple mouse haplotypes including a, b, d, j, k, s, and u. It does not bind to separated H-2 class I heavy chains or β2-m. Cell surface CD8 molecules bind to invariant sites of the H-2 heavy chains and can provide coreceptor signaling for Ag-mediated activation through the T cell receptor. H-2 class I molecules that present self-Ags can lead to self-tolerance of maturing CD8+ T cells due to thymic selection. Alternatively, these molecules can present foreign peptide Ags to mature peripheral CD8+ T cells resulting in cell-mediated immune responses against foreign Ags. H-2 class I Ags also serve as ligands for activating and inhibitory receptors expressed by NK cells and some T cells. The M1/42 antibody is useful for analyzing H-2 class I Ag expression on cells or cell lines. Cells from different experimental systems, eg, stressed cells that have undergone infection or transformation, may express little or no H-2 class I Ag. In contrast, cells undergoing activation or responding to certain factors (eg, interferons) may express upregulated levels of these Ags.
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 CompBead 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 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). 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. 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. CF™ is a trademark of Biotium, Inc.
10. BD Horizon Brilliant Ultraviolet 615 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.
