Detection of TLR6 in RAW 264.7 Mouse Cell Line by Flow Cytometry.
RAW 264.7 mouse monocyte/ macrophage cell line was stained with Rat Anti-Mouse TLR6 Monoclonal Antibody (Catalog # MAB1533, filled histogram) or isotype control antibody (Catalog # MAB006, open histogram), followed by Phycoerythrin-conjugated Anti-Rat IgG F(ab')2 Secondary Antibody (Catalog # F0105B).
Preparation and Storage
Reconstitute at 0.5 mg/mL in sterile PBS.
Reconstitution Buffer Available
The product is shipped at ambient temperature. Upon receipt, store it immediately at the temperature recommended below. *Small pack size (SP) is shipped with polar packs. Upon receipt, store it immediately at -20 to -70 °C
Stability & Storage
Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
12 months from date of receipt, -20 to -70 °C as supplied.
1 month, 2 to 8 °C under sterile conditions after reconstitution.
6 months, -20 to -70 °C under sterile conditions after reconstitution.
The Toll-like family of molecules are a group of integral membrane proteins that serve as pattern recognition receptors for microbial pathogens. There are at least eleven mouse and ten human members that activate the innate immune system following exposure to a variety of microbial species (1‑4). All Toll-like receptors (TLRs) are type I transmembrane (TM) proteins that exist either in the plasma membrane or in the membranes of endosomal structures (where they bind intracellular microbial nucleic acids). All TLRs also contain a large number of extracellular leucine-rich repeats (LRRs) and a cytoplasmic tail with a Toll/IL-1 receptor (TIR) domain. The mouse TLR6 cDNA encodes a 795 amino acid (aa) precursor that includes a 27 aa signal sequence, a 557 aa extracellular domain (ECD), a 21 aa transmembrane segment, and a 190 aa cytoplasmic domain. The ECD contains 14 Leu-rich repeats, and the cytoplasmic region contains one TIR domain (5). Within the ECD, mouse TLR6 shares 59% aa sequence identity with mouse TLR1 and 20‑27% aa sequence identitity with mouse TLR2, -3, -4, -5, -7, -8, -9, -11, -12, and -13. It shares 71%, 72%, and 86% aa sequence identity with bovine, human, and rat TLR6, respectively. TLR6 is expressed on the cell surface of macrophages, monocytes, neutrophils, and dermal endothelial cells in ligand-independent association with TLR2 (6‑9). TLR2 also associates with TLR1, a functional complex with specificity for distinct but related microbial ligands (6‑8). TLR6 and TLR2 cooperate in the recognition of acylated bacterial and mycoplasma lipopeptides, peptidoglycan, and glycosylphosphatidylinositols (7‑14). The cytoplasmic TIR domain is necessary and sufficient to initiate signal transduction which leads to activation of NF kappa B (7, 15).
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We have 1 review tested in 1 application: Flow Cytometry.
Bone marrow-derived macrophages,alveolar macrophages
Other Experimental Details
Other Experimental Details
Acute silica exposure reduced TLR 6 expression on F4-80+CD11c+ alveolar macrophages. C57Bl/6 wild-type mice were exposed to saline (25 μl, black line) or silica (1 mg, gray line) through intranasal aspiration. After 4 h, whole lungs were lavaged, cells immunostained, and surface markers assessed by flow symmetry using a BD FACS Aria II. Data shown as histograms relative to unstained control (dashed line).