Overview

  • Product name
    Anti-TLR9 antibody [5G5] (Biotin)
    See all TLR9 primary antibodies
  • Description
    Mouse monoclonal [5G5] to TLR9 (Biotin)
  • Host species
    Mouse
  • Conjugation
    Biotin
  • Tested applications
    Suitable for: IHC-P, ICC/IF, Flow Cyt, WB, IHC-Frmore details
  • Species reactivity
    Reacts with: Mouse, Dog
    Does not react with: Human
  • Immunogen

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

  • Positive control
    • RAW macrophages and TLR9 transfected HEK293 cells. RAW264.7 macrophages stimulated with IFN gamma.

Properties

Applications

Our Abpromise guarantee covers the use of ab17236 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Application Abreviews Notes
IHC-P Use at an assay dependent concentration.
ICC/IF Use at an assay dependent concentration.
Flow Cyt Use at an assay dependent concentration.

ab97679 - Mouse monoclonal IgG2a, is suitable for use as an isotype control with this antibody.

 

WB Use at an assay dependent concentration. Predicted molecular weight: 115 kDa.
IHC-Fr Use at an assay dependent concentration.

Target

  • Function
    Key component of innate and adaptive immunity. TLRs (Toll-like receptors) control host immune response against pathogens through recognition of molecular patterns specific of microorganisms. TLR9 is a nucleotide-sensing TLR which is activated by unmethylated cytidine-phosphate-guanosine (CpG) dinucleotides. Acts via MYD88 and TRAF6, leading to NF-kappa-B activation, cytokine secretion and the inflammatory response.
  • Tissue specificity
    Highly expressed in spleen, lymph node, tonsil and peripheral blood leukocytes, especially in plasmacytoid pre-dendritic cells. Levels are much lower in monocytes and CD11c+ immature dendritic cells. Also detected in lung and liver.
  • Sequence similarities
    Belongs to the Toll-like receptor family.
    Contains 26 LRR (leucine-rich) repeats.
    Contains 1 TIR domain.
  • Cellular localization
    Endoplasmic reticulum membrane. Endosome. Lysosome. Cytoplasmic vesicle > phagosome. Relocalizes from endoplasmic reticulum to endosome and lysosome upon stimulation with agonist.
  • Information by UniProt
  • Database links
  • Alternative names
    • CD 289 antibody
    • CD289 antibody
    • TLR 9 antibody
    • TLR9 antibody
    • TLR9_HUMAN antibody
    • Toll like receptor 9 antibody
    • Toll like receptor 9 isoform A precursor antibody
    • Toll like receptor 9 isoform B antibody
    • Toll-like receptor 9 antibody
    see all

Images

  • Lane 1 : Anti-TLR9 antibody [5G5] (Biotin) (ab17236) at 1/500 dilution
    Lane 2 : TLR9 Knockout

    All lanes : Mouse liver whole tissue lysate

    Lysates/proteins at 20 µg per lane.

    Developed using the ECL technique.

    Performed under reducing conditions.

    Predicted band size: 115 kDa


    Exposure time: 40 seconds


    Blocking agent: 5% Milk for 1 hour at 23°C.

    See Abreview

  • Immunfluorescence analysis of RAW264.7 cells, staining TLR9 with ab17236.

    Cells were untreated (Un) or treated with GO5 or GO100 for 24 hours, fixed and then permeabilized. Cells were incubated with primary antibody (1/100) for for 1 hour at 4°C in the dark. An AlexaFluor®488-conjugated goat anti-mouse IgG was used as the secondary antibody.
  • Flow cytometry analysis of RAW264.7 cells, staining TLR9 with ab17236.

    Cells were either untreated (green line) or treated (pink line) with GO5 or GO100 and fixed and permeabilized with 4% formaldehyde and 0.5% Tween-20. Cells were then incubated with primary antibody (1/100 dilution) for 1 hour at 4°C in the dark. An AlexaFluor®488-conjugated goat anti-mouse IgG was used as the secondary antibody.

References

This product has been referenced in:
  • Chen GY  et al. Simultaneous induction of autophagy and toll-like receptor signaling pathways by graphene oxide. Biomaterials 33:6559-69 (2012). ICC/IF, Flow Cyt ; Mouse . Read more (PubMed: 22704844) »
  • Chen GY  et al. Baculovirus transduction of mesenchymal stem cells triggers the toll-like receptor 3 pathway. J Virol 83:10548-56 (2009). Flow Cyt ; Human . Read more (PubMed: 19656899) »
See all 3 Publications for this product

Customer reviews and Q&As

1-4 of 4 Abreviews or Q&A

Application
Western blot
Sample
Mouse Tissue lysate - whole (liver)
Gel Running Conditions
Reduced Denaturing
Loading amount
20 µg
Treatment
TLR9 knockout
Specification
liver
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 23°C

Abcam user community

Verified customer

Submitted Aug 12 2015

Answer

Thank you for contacting us.

Unfortunately, all the references and images available are already on the datasheet. We are continuously updating our datasheets, and as soon as we gather further characterisation details about it, we will share it on the Website.

We also encourage our antibody’s users to send us the images obtained with the antibodies and earn Abpoints in compensation: https://www.abcam.com/abpoints.

However, all of our products are covered by our Abpromise® guarantee, which ensures that you can trust our products, and they should work in the tested species and applications stated on the datasheet, or we will offer a replacement, credit, or refund, if reported within 6 months of purchase.

To read about our Abpromise policy: https://www.abcam.com/index.html?pageconfig=abpromise

I hope this information is helpful to you. Please do not hesitate to contact us if you need any more advice or information.

Read More

Answer

Thank you for your e-mail regarding ab17236 - Mouse monoclonal [5G5] to TLR9 (Biotin). We are very sorry to hear the there are still problems with this product and have been discussing this complaint with some colleagues. We looked very carefully the information you provided to us. You mentioned that you tried staining 1x10E6 cells with between 0.1 and 0.5 ug/ml of primary antibody. What is the volume you actually used? We would advise you to use a concentration of 10 ug/ml ( 2 ug/200 ul). It seems that concentration of 0.1 ug/ml are very low. We use the following fixation and permeabilization protocol: - Wash the cells 2x with cold PBS. Temperature control at this point is of vital importance. Note: Cells should be kept on ice any time they are not spinning, and cells should be spun in a temperature controlled centrifuge at 4°C. - Fix the cells with 1% paraformaldehyde for 15 minutes at 4°C. Note: it is crucial to make sure that the cells are resuspended well. - Wash the cells 2x with ice cold PBS. - Permeabilize the cells with 100% ice cold methanol added drop wise while the cells are vortexing. It is crucial to make sure that the cells are resuspended thoroughly. Leave in methanol for 15 minutes. - Wash the cells 2x with cold PBS - Add approximately 1x10E6 cells to each FACS tube and wash with 1 ml of 0.1% saponin diluted in PBS with 2% fetal bovine serum added. - Resuspend cells in 1 ml of 0.1% saponin + 2% FBS and allow to sit for 30 minutes blocking at RT. - Add primary antibody We are not sure why you washed the cells between the permeabilization step and the primary antibody incubation. We would like to advise you to follow the above protocol (if this is not the protocol you normally use). I hope this information will be useful for you, please do not hesitate to contact Abcam if you still have further questions.

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Question
Answer

Thank you for your enquiry. This Ab is cited int following paper being used for WB: Ahmad-Nejad, P et al; Bacterial CpG-DNA and lipopolysaccharides activate Toll-like receptors at distinct cellular compartments. Eur J Immunol 2002, 32: 1958 Contained within this paper is a Western blot image pertaining to this product. If you need any further assistance then please get back in touch with us.

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Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"

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