Description: M9.D6 is generated against a peptide derived from the extracellular portion of mouse TLR9. Predomitly 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: This M9.D6 antibody has been reported for use in intracellular flow cytometric analysis, and immunoblotting (WB).(Fluorochrome conjugated M9.D6 is recommended for use in intracellular flow cytometry.). Applications Tested: The M9.D6 antibody has been tested by intracellular flow cytometric analysis of fixed and permeabilized mouse TLR9 transfected cells. This can 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. Purity: Greater than 90%, as determined by SDS-PAGE. Aggregation: Less than 10%, as determined by HPLC. Filtration: 0.2 µm post-manufacturing filtered. The protein encoded by this gene is a member of the Toll-like receptor family which plays a fundamental role in pathogen recognition and activation of innate immunity. TLRs are highly conserved from Drosophila to humans and share structural and functional similarities. They recognize pathogen-associated molecular patterns that are expressed on infectious agents, and mediate the production of cytokines necessary for the development of effective immunity. The various TLRs exhibit different patterns of expression. This gene is preferentially expressed in immune cell rich tissues, such as spleen, lymph node, bone marrow and peripheral blood leukocytes. Studies in mice and human indicate that this receptor mediates cellular response to unmethylated CpG dinucleotides in bacterial DNA to mount an innate immune response.
|Flow Cytometry, Western Blot|
|PBS with 0.09% sodium azide; pH 7.2|
For Research Use Only.
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