Product Information
Contents: Fluorescein isothiocyanate (FITC) anti-mouse Toll-like receptor 9 (CD289, TLR9, TLR-9)
Catalog Number: 11-9093
Sizes: 25 ug
Formulation: Phosphate buffer pH 7.2, 150 mM NaCl, 0.09% NaN3
Storage Conditions: Store at 4°C. DO NOT FREEZE. LIGHT-SENSITIVE MATERIAL.
Clone: M9.D6
Isotype: Rat IgG2a
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Staining of fixed and permeabilized non-transfected (left) and mouse TLR9 cDNA transiently transfected (right) 293T cells with 0.25 μg FITC Rat IgG2a Iso Cntrl (cat. 11-4321) (open histogram) or 0.5 μg FITC anti-mouse TLR9 (M9.D6) (colored histogram). Total viable cells were used for analysis.
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Description
M9.D6 is generated against a peptide derived from the extracellular portion of mouse TLR9. Predominantly expressed as an intracellular protein, TLR9 is a ~115-120 kDa molecule which mediates response to unmethylated CpG dinucleotides in bacterial DNA. CpG DNA induces a strong T-helper-1-like inflammatory response and the proliferation of TLR9+ B cells. When stimulated with CpG DNA, TLR9-deficient (TLR9-/-) mice lacked splenocyte proliferation, inflammatory cytokine production from macrophages, and dendritic cell maturation, as compared with normal mice. To date, at least ten members of the Toll family have been identified. This family of type I transmembrane proteins is characterized by an extracellular domain with leucine-rich repeats and a cytoplasmic domain with homology to the type I IL-1 receptor. Members of the TLR family are involved in recognition and response to different microbial components including lipoproteins, peptidoglycans, and nucleic acids and play important roles in innate immunity and inflammation. TLR9 is not detected by flow cytometry using this antibody on RBC-lysed mouse splenocytes stained for intracellular TLR9. This may be due to limitations of antigen detection by flow cytometry. Further studies are needed to determine the relationship between mRNA expression and protein detection.
Applications Reported
For research use only, not for diagnostic or therapeutic use. This M9.D6 antibody has been reported for use in intracellular flow cytometric analysis.
Applications Tested
This M9.D6 antibody has been tested by intracellular staining and flow cytometric analysis of fixed and permeabilized mouse TLR9 transfected 293T cells. This can be used at less than or equal to 0.5 μg per million cells in a 100 μl total staining volume. It is recommended that the antibody be carefully titrated for optimal performance in the assay of interest.
References
Ahmad-Nejad P, Hacker H, Rutz M, Bauer S, Vabulas RM, Wagner H. 2002. Bacterial CpG-DNA and lipopolysaccharides activate Toll-like receptors at distinct cellular compartments. Eur J Immunol. 32(7): 1958-68.
Fukata M, Breglio K, et al. 2008. The Myeloid Differentiation Factor 88 (MyD88) Is Required for CD4+ T Cell Effector Function in a Murine Model of Inflammatory Bowel Disease. J Immunol. 180(3):1886-94. (M9.D6, IC Flow,
PubMed)
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