MERTK Monoclonal Antibody (DS5MMER), Super Bright™ 600, eBioscience™, Invitrogen™ 

Description

Description: This DS5MMER monoclonal antibody recognizes mouse MerTK, a 170-210 kDa member of the TAM family of tyrosine kinase receptors that also includes Axl and Tyro3. MerTK is expressed on tissue macrophages and is involved in the removal of apoptotic cells. This process relies on two soluble ligands of MerTK, Protein S and Gas6 that bind to phosphatidylserine found on the outer leaflet of the plasma membrane of cells undergoing apoptosis. Upon binding these ligands, MerTK undergoes autophosphorylation at multiple tyrosine residues that activate the PI3K and Akt pathways. This results in the phagocytosis of apoptotic cells and also results in the direct inhibition of TLR-induced production of pro-inflammatory cytokines. In addition, MerTK may function as a putative entry receptor for filoviruses. Deficiency of MerTK causes general autoimmunity, inflammation and accumulation of apoptotic bodies. MerTK is constitutively released from the cell surface by metalloproteinases and thus is present in the serum and culture medium. This process can be enhanced by stimulation with LPS. MerTK can be used to help discriminate macrophages from dendritic cells. MerTK is often expressed on maligt cells and may be implicated in immune evasion. The DS5MMER antibody will work on paraformaldehyde-fixed cells. Applications Reported: This DS5MMER antibody has been reported for use in flow cytometric analysis.

Applications Tested: This DS5MMER antibody has been tested by flow cytometric analysis of mouse resident peritoneal cells. This can be used at less than or equal to 1 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 Complete Staining Buffer (Product No. SB-4401) 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 specificperformance should be determined empirically. Excitation: 405 nm; Emission: 600 nm; Laser: Violet Laser Super Bright Polymer Dyes are sold under license from Becton, Dickinson and Company. Receptor tyrosine kinase that transduces signals from the extracellular matrix into the cytoplasm by binding to several ligands including LGALS3, TUB, TULP1 or GAS6. Regulates many physiological processes including cell survival, migration, differentiation, and phagocytosis of apoptotic cells (efferocytosis). Ligand binding at the cell surface induces autophosphorylation of MERTK on its intracellular domain that provides docking sites for downstream signaling molecules. Following activation by ligand, interacts with GRB2 or PLCG2 and induces phosphorylation of MAPK1, MAPK2, FAK/PTK2 or RAC1. MERTK signaling plays a role in various processes such as macrophage clearance of apoptotic cells, platelet aggregation, cytoskeleton reorganization and engulfment. Functions in the retinal pigment epithelium (RPE) as a regulator of rod outer segments fragments phagocytosis. Plays also an important role in inhibition of Toll-like receptors (TLRs)-mediated innate immune response by activating STAT1, which selectively induces production of suppressors of cytokine signaling SOCS1 and SOCS3.