Description: The BM8 monoclonal antibody reacts with mouse F4/80 antigen, an approximately 160 kDa surface receptor. It belongs to the EGF-TM7 family of proteins. As such it contains seven EGF-like domains on its extracellular N-terminus, seven transmembrane spanning sequences, and an intracellular C-terminal domain showing homology to other TM7 superfamily members. The F4/80 antigen is expressed by a majority of mature macrophages and is one of the best markers for this population of cells. However, other cell types, such as peritoneal eosinophils, Langerhans cells, and some other dendritic cell subtypes, have been reported to express this antigen as well. Expression of F4/80 commences during early myeloid development in vivo and can be upregulated on BM cells stimulated in vitro with M-CSF. Some populations of macrophages, especially in the lymphoid microenvironment, may be devoid of F4/80. Applications Reported: This BM8 antibody has been reported for use in flow cytometric analysis. Applications Tested: This BM8 antibody has been tested by flow cytometric analysis of resident peritoneal macrophages. This may be used at less than or equal to 0.5 μg per test. A test is defined as the amount (μg) of antibody that will stain a cell sample in a final volume of 100 μL. Cell number should be determined empirically but can range from 10^5 to 10^8 cells/test.It is recommended that the antibody be carefully titrated for optimal performance in the assay of interest. Super Bright 600 is a tandem dye that can be excited with the violet laser line (405 nm) and emits at 600 nm. We recommend using a 610/20 bandpass filter. Please make sure that your instrument is capable of detecting this fluorochrome. When using two or more Super Bright dye-conjugated antibodies in a staining panel, it is recommended to use Super Bright Staining Buffer (Product No. SB-4400) to minimize any non-specific polymer interactions. Please refer to the datasheet for Super Bright Staining Buffer for more information. Light sensitivity: This tandem dye is sensitive to photo-induced oxidation. Please protect this vial and stained samples from light. Fixation: Samples can be stored in IC Fixation Buffer (Product No. 00-8222) (100 μL of cell sample + 100 μL of IC Fixation Buffer) or 1-step Fix/Lyse Solution (Product No. 00-5333) for up to 3 days in the dark at 4°C with minimal impact on brightness and FRET efficiency/compensation. Some generalizations regarding fluorophore performance after fixation can be made, but clone specific performance should be determined empirically. Excitation: 405 nm; Emission: 600 nm; Laser: Violet Laser F4/80 antigen is a 160 kDa cell surface glycoprotein that is a member of the EGF TM7 family of proteins and shares 68% overall amino acid identity with human EGF module containing mucin like hormone receptor 1 (EMR1). Expression of F4/80 is heterogeneous and is reported to vary during macrophage maturation and activation. The F4/80 antigen is expressed on a wide range of mature tissue macrophages including Kupffer cells, Langerhans, microglia, macrophages located in the gut lamina propria, peritoneal cavity, lung, thymus, bone marrow stroma and macrophages in the red pulp of the spleen. F4/80 expression has also been reported on a subpopulation of dendritic cells but is absent from macrophages located in T cell areas of the spleen and lymph node. The ligands and biological functions of the F4/80 antigen have not yet been determined but recent studies suggest a role for F4/80 in the generation of efferent CD8+ve regulatory T cells.
|4° C, store in dark, DO NOT FREEZE!|
|Super Bright 600|
|PBS with BSA and 0.09% sodium azide; pH 7.2|