Ba/F3 cell line transfected with mouse TLR4 /MD2 complex
Formulation
0.2 µm filtered in phosphate-buffered solution, pH 7.2, containing no preservative. Endotoxin level is <0.01 EU/µg of the protein (<0.001 ng/µg of the protein) as determined by the LAL test.
Preparation
The Ultra-LEAF™ (Low Endotoxin, Azide-Free) antibody was purified by affinity chromatography.
Concentration
1mg/ml
Recommended Usage
Each lot of this antibody is quality control tested by immunofluorescent staining with flow cytometric analysis. For immunofluorescent staining, the suggested use of this reagent is ≤0.25 µg per million cells in 100 µl volume or 100 µl of whole blood. It is recommended that the reagent be titrated for optimal performance for each application.
Applications
FC - Quality tested IHC - Validated ICFC, IF, IP, Block - Reported in the literature
Application Notes
In addition to flow cytometric uses, the MTS510 antibody has also been used for immunoprecipitation, immunofluorescence, and blocking of LPS-induced stimulation. The LEAF™ purified antibody (Endotoxin <0.1 EU/μg, Azide-Free, 0.2 μm filtered) is recommended for functional assays (Cat. No. 117607). For highly sensitive assays, we recommend Ultra-LEAF™ purified antibody (Cat. No. 117612) with a lower endotoxin limit than standard LEAF™ purified antibodies (Endotoxin <0.01 EU/µg).
Application References
1. Akashi S, et al. 2000. J. Immunol. 164:3471. (FC, ICFC, IP, Block) 2. Kawasaki K, et al. 2003. J. Immunol. 170:413. (FC) 3. Nalbandian G, et al. 2005. J. Immunol. 175:2666. (FC) PubMed 4. Kanada S, et al. 2011. Blood 117:2211. PubMed 5. Laplate P, et al. 2011. J. Biol Chem. 286:42494. PubMed 6. Khan MA, et al. 2006. Infect. Immun. 74:2522. (IF)
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Description
Toll-like receptors are highly conserved from Drosophila to humans and share structural and functional similarities. They are type I transmembrane signaling receptors which activate the innate immune system in response to pathogen invasion. So far, at least 13 TLR members have been identified. The secretory protein, MD2, associates with cell-surface bound TLR4 (CD284) on monocytes, B cells and T cells. TLR4 has been recognized as critical for host recognition of bacterial LPS in conjunction with MD2.
Storage
The antibody solution should be stored undiluted at 4°C. This Ultra-LEAF™ solution contains no preservative; handle under aseptic conditions.