MERTK Monoclonal Antibody (DS5MMER), Super Bright™ 780, eBioscience™
Isotype: IgG2a, kappa
Format: Super Bright™ 780
Application: Flow Cytometry
Tested Dilution: 1.0 µg/test
Concentration: 0.2 mg/mL
Storage: 4° C, store in dark, DO NOT FREEZE!
Formulation: PBS, pH 7.2, containing 0.09% sodium azide
Purification: Affinity chromatography
Data Sheet: TDS
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 malignant 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 resident peritoneal macrophages. This may 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 780 is a tandem dye that can be excited with the violet laser line (405 nm) and emits at 780 nm. We recommend using a 780/60 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 # SB-4401) to minimize any non-specific polymer interactions. Please refer to the datasheet for Super Bright Staining Buffer for more information.
In some experiments, we have observed that compensation values for Super Bright 780-conjugated antibodies are higher in the violet 450/50 channel when using UltraComp eBeads microspheres (Product # 01-2222-42) as compared to single-color stained cells. In such circumstances, we would recommend setting compensation with cells. We have also observed this in some experiments using AbC Total Antibody Compensation beads (Product # A10497).
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 # 00-8222) (100 µL of cell sample + 100 µL of IC Fixation Buffer) or 1-step Fix/Lyse Solution (Product # 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: 780 nm; Laser: Violet Laser
Super Bright Polymer Dyes are sold under license from Becton, Dickinson and Company.
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.