MERTK Monoclonal Antibody (DS5MMER), Super Bright™ 702, eBioscience™, Invitrogen™

Description

Description: This JAV4 monoclonal antibody recognizes human V-Set and Immunoglobulin domain containing 4 (VSIG4), also known as Complement Receptor of the Immunoglobulin superfamily (CRIg) or Z39Ig. There are two variants of human VSIG4 that result from alternative splicing. Isoform 1 is longer and more prevalent than isoform 2, and this JAV4 antibody recognizes both. VSIG4 is a type I transmembrane glycoprotein structurally related to the B7 family of immune regulatory proteins. It contains one complete V-type Ig domain and one truncated C-type Ig domain. VSIG4 is exclusively expressed on tissue resident and tumor infiltrating macrophages. It has been shown to bind complement components C3b and iC3b. This binding inhibits the alternative complement pathway and facilitates phagocytosis of complement-opsonized pathogens. VSIG4 has also been reported to suppress T cell activation, proliferation and IL-2 production thereby playing a role in the maintece of peripheral T cell tolerance and suppression of established inflammation. Expression of VSIG4 on tumor-infiltrating macrophages suggests its role in immune evasion. Pro-inflammatory stimuli such as TNF and LPS have been reported to down-regulate the expression of VSIG4. The JAV4 antibody will recognize VSIG4 on cells that have been formaldehyde-fixed and permeabilized. This antibody does not cross-react with mouse or rat VSIG4. Applications Reported: This JAV4 antibody has been reported for use in flow cytometric analysis.

Applications Tested: This DS5MMER antibody has been tested by flow cytometric analysis of resident peritoneal macrophages. This may be used at less than or equal to 1.0 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 702 is a tandem dye that can be excited with the violet laser line (405 nm) and emits at 702 nm. We recommend using a 710/50 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: 702 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.