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TLR9 Antibody (5G5)

Novus Biologicals, part of Bio-Techne | Catalog # NBP1-51156

Novus Biologicals, part of Bio-Techne
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NBP1-51156

Key Product Details

Species Reactivity

Human, Mouse, Canine

Applications

Flow Cytometry, Immunocytochemistry/ Immunofluorescence, Immunohistochemistry, Immunohistochemistry-Frozen, Immunohistochemistry-Paraffin, Western Blot

Label

Unconjugated

Antibody Source

Monoclonal Mouse IgG2A Clone # 5G5

Concentration

0.1 mg/ml

Product Specifications

Immunogen

Purified fusion protein of extracellular domain of human TLR9 (AA 1-815) and human IgGFc

Reactivity Notes

Please note that this antibody is reactive to Mouse and derived from the same host, Mouse. Additional Mouse on Mouse blocking steps may be required for IHC and ICC experiments. Please contact Technical Support for more information.

Clonality

Monoclonal

Host

Mouse

Isotype

IgG2A

Description

Positive Control: RAW264.7 macrophages stimulated with IFNg

Scientific Data Images for TLR9 Antibody (5G5)

Flow Cytometry: TLR9 Antibody (5G5) [NBP1-51156]

Flow Cytometry: TLR9 Antibody (5G5) [NBP1-51156]

Flow Cytometry: TLR9 Antibody (5G5) [NBP1-51156] - THP1 cells were incubated with IgG2a isptype controle Cells (140000) were permeabilized with saponin and stained with 0.4 ug 5G5
Flow Cytometry: TLR9 Antibody (5G5) [NBP1-51156]

Flow Cytometry: TLR9 Antibody (5G5) [NBP1-51156]

Flow Cytometry: TLR9 Antibody (5G5) [NBP1-51156] - THP1 cells were incubated with IgG2a isptype controle (A) or a-TLR9 5G5 mAb

Applications for TLR9 Antibody (5G5)

Application
Recommended Usage

Flow Cytometry

1:50

Immunocytochemistry/ Immunofluorescence

4ug/400ul

Immunohistochemistry

1:10-1:500

Immunohistochemistry-Frozen

1:100

Immunohistochemistry-Paraffin

1:50

Western Blot

2 ug/ml
Application Notes
IHC-Frozen: 10um sections were fixed with acetone for 10 minutes. PBS washed sections were incubated with 5G5 1:100 in 1% BSA for 30 minutes at RT. IF: cells were fixed with 2% formalin for 15 minutes at RT and permeabilized with a mAb (4ug/400ul) containing buffer (PBS, 0.2% BSA, 0.2% saponin) for 1 hour. FC: RAW264.7 cells were fixed for 15 minutes with 4% formalin and permeabilized (PBS, 0.5%BSA, 0,5% saponin) at RT. IHC-Paraffin: paraffin embedded tissues 5um sections were made. After antigen retrieval (0.01mol/l, pH6 sodium citrate) and quenching of endogenous peroxidase, sections were blocked with 0.5% ovalbumin and 0.1% gelatin for 20 minutes at RT. Sections were incubated with 5G5 for 1 hour at 37C. WB: reduced lysates were resolved by 10% SDS-PAGE and blotted on nitrocellulose. After blocking with 5% skimmed milk TLR9 was detected with 2 ug/ml 5G5

Formulation, Preparation, and Storage

Purification

Protein A or G purified

Formulation

PBS, containing 0.1% BSA

Preservative

0.02% Sodium Azide

Concentration

0.1 mg/ml

Shipping

The product is shipped with polar packs. Upon receipt, store it immediately at the temperature recommended below.

Stability & Storage

Store at 4C.

Background: TLR9

The monoclonal antibody 5G5 recognizes human Toll-like receptor 9. Toll-like receptors (TLRs) are highly conserved from Drosophila to humans and share structural and functional similarities. TLRs constitute of a family of pattern recognition receptors (PRRs) that mediate cellular responses to a large variety of pathogens (viruses, bacteria, and parasites) by specific recognition of so-called pathogen-associated molecular patterns. Activation of TLRs, a family of at least 11 different members that function either as homo- or heterodimers, leads to activation of NF'B-dependent and IFN-regulatory factor-dependent signaling pathways. TLRs have a central role in innate immunity and are also required for the development of an adaptive immune response. TLRs are expressed by various cells of the immune system, such as macrophages and dendritic cells. They recognize and respond to molecules derived from bacterial, viral and fungal pathogens. Whereas most TLRs are expressed on the cell surface, TLR9 is expressed intracellularly within one or more endosomal compartments and recognizes nucleic acids. TLR9 detects a rather subtle difference in the DNA of vertebrates compared with that of pathogens. Vertebrate genomic DNAs have mostly methylated CpG dinucleotides where bacterial and viral DNAs have unmethylated CpG dinucleotides. TLR9 undergoes relocation from endoplasmic reticulum to CpG-ODN-containing endosomes. In these endosomes TLR9 becomes a functional receptor after proteolytic cleavage. TLR9 exists as a preformed homodimer and CpG-ODN binding promotes its conformational change, bringing the cytoplasmic TIR-like domains close to each other. This allows a recruitment of the key adapter protein MyD88 which initiates a signalling cascade. The only human immune cell types known to constitutively express TLR9 and to be activated by CpG ODN are pDCs and B cells. TLR9 triggering induces an activation phenotype in the B cells and pDCs, characterized by the expression of costimulatory molecules, resistance to apoptosis, and induces Th1-type immune response profiles.

Long Name

Toll-like Receptor 9

Alternate Names

CD289

Gene Symbol

TLR9

Additional TLR9 Products

Product Documents for TLR9 Antibody (5G5)

Certificate of Analysis

To download a Certificate of Analysis, please enter a lot number in the search box below.

Product Specific Notices for TLR9 Antibody (5G5)

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Primary Antibodies are guaranteed for 1 year from date of receipt.

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