TLR9 Antibody (5G5)

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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] - THP1 cells were incubated with IgG2a isptype controle (A) or a-TLR9 5G5 mAb

Product Details

Summary
Reactivity Mu, CaSpecies Glossary
Applications WB, Flow, ICC/IF, IHC, IHC-Fr, IHC-P
Clone
5G5
Clonality
Monoclonal
Host
Mouse
Conjugate
Unconjugated
Concentration
0.1 mg/ml

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TLR9 Antibody (5G5) Summary

Immunogen
Purified fusion protein of extracellular domain of human TLR9 (AA 1-815) and human IgGFc
Isotype
IgG2a
Clonality
Monoclonal
Host
Mouse
Gene
TLR9
Purity
Protein A or G purified
Innovator's Reward
Test in a species/application not listed above to receive a full credit towards a future purchase.

Applications/Dilutions

Dilutions
  • Western Blot 2 ug/ml
  • Flow Cytometry 1:50
  • Immunocytochemistry/Immunofluorescence 4ug/400ul
  • Immunohistochemistry 1:10-1:500
  • Immunohistochemistry-Frozen 1:100
  • Immunohistochemistry-Paraffin 1:50
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
Agonist

Reactivity Notes

Cross reacts with Canine and Mouse. 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.

Packaging, Storage & Formulations

Storage
Store at 4C. Do not freeze.
Buffer
PBS, containing 0.1% BSA
Preservative
0.02% Sodium Azide
Concentration
0.1 mg/ml
Purity
Protein A or G purified

Notes

Positive Control: RAW264.7 macrophages stimulated with IFNg

Alternate Names for TLR9 Antibody (5G5)

  • CD289 antigen
  • CD289
  • TLR9
  • toll-like receptor 9

Background

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.

Limitations

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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Product General Protocols

Video Protocols

WB Video Protocol
ICC/IF Video Protocol

FAQs for TLR9 Antibody (NBP1-51156) (0)

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Secondary Antibodies

 

Isotype Controls

Other Available Formats

Biotin Labeled NB600-1420

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Topics in CD11b: The innate immune response
Integrins are transmembrane receptors composed of alpha and beta chains, where beta-integrins are mainly expressed in leukocytes. Leukocytes are white blood cells that act in the immune system to defend our body against foreign pathogens.  Integrin...  Read full blog post.

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MHC molecules (also known as major histocompatibility complex molecules) assist in the presentation of antigens to T cells in order to eradicate foreign pathogens.  These molecules are highly polymorphic, meaning that they exist in multiple varian...  Read full blog post.

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Toll like receptors (TLRs) are highly conserved proteins that are first known for their role in pathogen recognition and immune response activation.  In order to elicit the necessary immune response in reaction to a foreign pathogen, TLRs trigger cy...  Read full blog post.

IRAK4: The "master IRAK" critical for initiating immune responses
IRAK4, also known as Interleukin-1 receptor-associated kinase 4, is a serine/threonine-protein kinase that plays a critical role in initiating innate and adaptive immune responses against foreign pathogens. It activates NF-kappaB in both Toll-like rec...  Read full blog post.

TLR9: For Whom the Cell Tolls
The Toll-like receptor 9 (TLR9) protein, also known as CD289, belongs to the family of Toll-like receptor (TLR) proteins which play a large role in pathogen recognition and the activation of innate immunity. Scientists using TLR9 antibodies have found...  Read full blog post.

TLR9: Tollgate to Immunity
Toll-like receptors (TLRs) play an essential role in the activation of innate immunity, and TLRs are expressed in a large number of immune cells as well as in epithelial cells. TLR9 recognizes synthetic oligodeoxynucleotides (ODN) containing unmethyla...  Read full blog post.

TLR9, Infectious Disease and Cancer
Toll-like receptor 9 (TLR9) is a protein encoded by TLR9 gene in humans. It is also known as cluster of differentiation 289 (CD289) and is a member of TLR family. Proteins from TLR family are transmembrane proteins that expressed in both antigen-resen...  Read full blog post.

TLR9 Antibodies in Immunity Research
Toll-like receptor 9 (TLR9) is a member of the toll-like receptor family that plays a key role in pathogen recognition and activation of innate immunity. Scientists using TLR9 antibodies have found the protein is highly conserved from Drosophila to hu...  Read full blog post.

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Bioinformatics

Gene Symbol TLR9
Entrez