Description
The MAR-1 monoclonal antibody specifically recognizes Fc-epsilon RI-alpha (FceR1 alpha, also known as FcεR1α or FcER1a) which is likewise known as the IgE Fc receptor subunit alpha. FcεR1α is a type I transmembrane glycoprotein that is encoded by Fcer1a (Fc receptor, IgE, high affinity I, alpha polypeptide) which belongs to the Ig gene superfamily. As a single IgE-binding alpha subunit, FcεR1α complexes with signal transducing subunits including one beta subunit (FcεRIβ encoded by Ms4a2) and two disulfide-linked gamma subunits (FcεRIγ encoded by Fcer1g) to form the high-affinity receptor for IgE, Fc epsilon RI (FcεR1 or FcER1). FcεR1 is expressed on basophils and mast cells. Binding of cognate antigens (allergens) to FcεR1 with bound IgE antibodies leads to cellular activation and the release of mediators, including histamine and cytokines, that are responsible for allergic reactions. Tang et al. (2019) have reported that the MAR-1 antibody crossreacts with two other mouse Fc receptor chains, FcγRI (CD64) and FcγRIV (CD16-2), that are expressed by monocytes, macrophages, dendritic cells, or neutrophils. They suggested MAR-1 binding is specific for FcεRIa only on mast cells and basophils and that for other cell types reactive with the MAR-1 antibody, it may be appropriate to refer to these as MAR-1-positive (MAR-1+) cells.
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. 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. Please observe the following precautions: We recommend that special precautions be taken (such as wrapping vials, tubes, or racks in aluminum foil) to protect exposure of conjugated reagents, including cells stained with those reagents, to any room illumination. Absorption of visible light can significantly affect the emission spectra and quantum yield of tandem fluorochrome conjugates.
8. For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
9. An isotype control should be used at the same concentration as the antibody of interest.
10. CF™ is a trademark of Biotium, Inc.
11. Tandem fluorochromes contain both an energy donor and an energy acceptor. Although every effort is made to minimize the lot-to-lot variation in the efficiency of the fluorochrome energy transfer, differences in the residual emission from the donor may be observed. Additionally, multi-laser cytometers may directly excite both the donor and acceptor fluorochromes. Therefore, we recommend for every tandem conjugate, a matched individual single-stain control be acquired for generating a compensation or spectral unmixing matrix.
