Product Information

Contents: Cy5 anti-human Toll-like receptor 9 (CD289, TLR9, TLR-9)
Catalog Number: 19-9099
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: eB72-1665
Isotype: Rat IgG2a, κ

Intracellular staining of non-transfected (left) and human TLR9 transfected (right) HEK293 cells with PE Rat IgG2a isotype control (open histogram) or PE anti-human TLR9 (eB72-1665) (blue histogram).

Available Formats of This Product
Cat. No.	Format	Excite (nm)	Emit (nm)	Reported Applications
14-9099	Affinity Purified anti-human Toll-like receptor 9 (CD289, TLR9, TLR-9)	N/A	N/A	IC Flow IHC IHC(Paraffin) IP WB
12-9099	PE anti-human Toll-like receptor 9 (CD289, TLR9, TLR-9)	488	575	IC Flow

Questions? Please consult our answers to frequently asked questions at http://www.ebioscience.com/faq.

Description

eB72-1665 is generated against a portion of human toll-like receptor 9 (aa 273-288), a molecule reported to be expressed predominantly intracellularly. 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-positive human 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 twelve 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 lysed whole human blood and/or isolated human PBMC stained for cell surface or intracellular TLR9. This may be due to limitations of antigen detection by flow cytometry. Human pDCs matured in the presence of IL-3 have been reported to stain with eB72-1665 by immunofluorescence microscopy (Nat Immunol. 5:190). Human Epithelial Cell lines were also reported to stain with this mAb (J. Immunol. 173: 1219). Further studies are needed to determine the relationship between mRNA expression and protein detection by flow cytometry.

Applications Reported

For research use only, not for diagnostic or therapeutic use. The eB72-1665 antibody has been reported for use in intracellular flow cytometric analysis of fixed and permeabilized cells. Further studies are under way with this monoclonal antibody to define the expression pattern of TLR9 on normal human blood leukocytes.

Applications Tested

The eB72-1665 antibody has been tested by intracellular flow cytometric analysis of hTLR9 transfected and non-transfected cells. This can be used at less than or equal to 1 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

Latz E, Schoenemeyer A, Visintin A, Fitzgerald KA, Monks BG, Knetter CF, Lien E, Nilsen NJ, Espevik T, Golenbock DT. 2004. TLR9 signals after translocating from the ER to CpG DNA in the lysosome. Nat Immunol. 5(2):190-8.

Juliane Platz, Christoph Beisswenger, Alexander Dalpke, Rembert Koczulla, Olaf Pinkenburg, Claus Vogelmeier, and Robert Bals. 2004. Microbial DNA Induces a Host Defense Reaction of Human Respiratory Epithelial Cells. J. Immunol. 173: 1219 - 1223.

Leifer, CA, Kennedy, MN, Mazzoni, A, Lee, CW, Kruhlak, MJ, Segal, DM. 2004. TLR9 is Localized in the Endoplasmic Reticulum Prior to Stimulation. J Immun. 173:1179-1183.

Jozsef L, Khreiss T, El Kebir D, Filep JG. Activation of TLR-9 induces IL-8 secretion through peroxynitrite signaling in human neutrophils. J Immunol. 2006 Jan 15;176(2):1195-202. (eB72-1665, FC, PubMed)

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FOR NON-COMMERCIAL RESEARCH USE ONLY. NOT FOR THERAPEUTIC OR IN VIVO APLICATIONS. OTHER USE NEEDS LICENSE FROM AMERSHAM PHARMACIA BIOTECH INC. UNDER U.S. PATENT Nos. 4,981,977 AND 5,268,486 AND OTHER PATENTS PENDING.

THIS MATERIAL IS SUBJECT TO PROPRIETARY RIGHTS OF AMERSHAM PHARMACIA BIOTECH INC. AND CARNEGIE MELLON UNIVERSITY AND MADE AND SOLD UNDER LICENSE FROM AMERSHAM PHARMACIA BIOTECH INC.  THIS PRODUCT IS LICENSED FOR SALE ONLY FOR RESEARCH.   IT IS NOT LICENSED FOR ANY OTHER USE.    THERE IS NO IMPLIED LICENSE HEREUNDER FOR ANY COMMERCIAL USE.   COMMERCIAL USE shall include:

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