Overview

  • Product name
    Anti-TLR2 antibody [T2.5] (FITC)
    See all TLR2 primary antibodies
  • Description
    Mouse monoclonal [T2.5] to TLR2 (FITC)
  • Host species
    Mouse
  • Conjugation
    FITC. Ex: 493nm, Em: 528nm
  • Tested applications
    Suitable for: IHC-Fr, Flow Cyt, IP, ICC/IF, Functional Studiesmore details
  • Species reactivity
    Reacts with: Mouse, Human
  • Immunogen

    Mouse TLR2 peptide.

  • Positive control
    • RAW264.7 cells.

Properties

Applications

Our Abpromise guarantee covers the use of ab59711 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-Fr 1/50.
Flow Cyt Use 10µl for 106 cells.

ab91356 - Mouse monoclonal IgG1, is suitable for use as an isotype control with this antibody.

IP 1/50.
ICC/IF 1/50.
Functional Studies Use at an assay dependent concentration.

Target

  • Function
    Cooperates with LY96 to mediate the innate immune response to bacterial lipoproteins and other microbial cell wall components. Cooperates with TLR1 to mediate the innate immune response to bacterial lipoproteins or lipopeptides. Acts via MYD88 and TRAF6, leading to NF-kappa-B activation, cytokine secretion and the inflammatory response. May also promote apoptosis in response to lipoproteins. Recognizes mycoplasmal macrophage-activating lipopeptide-2kD (MALP-2), soluble tuberculosis factor (STF), phenol-soluble modulin (PSM) and B.burgdorferi outer surface protein A lipoprotein (OspA-L) cooperatively with TLR6.
  • Tissue specificity
    Highly expressed in peripheral blood leukocytes, in particular in monocytes, in bone marrow, lymph node and in spleen. Also detected in lung and in fetal liver. Levels are low in other tissues.
  • Sequence similarities
    Belongs to the Toll-like receptor family.
    Contains 14 LRR (leucine-rich) repeats.
    Contains 1 TIR domain.
  • Post-translational
    modifications
    Glycosylation of Asn-442 is critical for secretion of the N-terminal ectodomain of TLR2.
  • Cellular localization
    Membrane.
  • Information by UniProt
  • Database links
  • Alternative names
    • CD282 antibody
    • CD282 antigen antibody
    • TIL 4 antibody
    • TIL4 antibody
    • TLR 2 antibody
    • TLR2 antibody
    • TLR2_HUMAN antibody
    • Toll like receptor 2 antibody
    • Toll like receptor 2 precursor antibody
    • Toll-like receptor 2 antibody
    • Toll/interleukin 1 receptor like 4 antibody
    • Toll/interleukin 1 receptor like protein 4 antibody
    • Toll/interleukin receptor like protein 4 antibody
    • Toll/interleukin-1 receptor-like protein 4 antibody
    see all

Images

  • Overlay histogram showing THP1 cells stained with ab59711 (red line). The cells were fixed with 80% methanol (5 min) and incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab59711, 1µg/1x106 cells) for 30 min at 22ºC. Isotype control antibody (black line) was mouse IgG1 FITC (1µg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed. This antibody gave a positive signal in THP1 cells fixed with 4% paraformaldehyde used under the same conditions.

    Please note that Abcam do not have any data for use of this antibody on non-fixed cells. We welcome any customer feedback.

References

This product has been referenced in:
  • Moretti IF  et al. Plasmatic membrane toll-like receptor expressions in human astrocytomas. PLoS One 13:e0199211 (2018). Read more (PubMed: 29912993) »
  • Sun W  et al. Crosstalk between TLR2 and Sphk1 in microglia in the cerebral ischemia/reperfusion-induced inflammatory response. Int J Mol Med 40:1750-1758 (2017). Read more (PubMed: 29039449) »
See all 6 Publications for this product

Customer reviews and Q&As

1-3 of 3 Abreviews or Q&A

Question
Answer

Thank you for your phone call.

The free of charge replacement antibody you requested, ab24192, will ship on order ***. This antibody has not been validated for use on frozen sections, but may be suitable for that purpose. The antibody may require antigen retrieval for longer PFA fixations. If you are unable to use a shorter fixation method, the attached reference provides details about how to perform heat-mediated antigen retrieval on frozen tissue sections. I hope this helps, please let me know if you need any additional information or assistance.

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Question
Answer

I have found another reference that is more encouraging:

TLR2 is constitutively expressed within the kidney and participates in ischemic renal injury through both MyD88-dependent and -independent pathways.
J Immunol. 2007 May 15;178(10):6252-8.

http://www.jimmunol.org/content/178/10/6252.full

The mouse tissue was PFA-fixed and embedded frozen in OCT, and it does not appear any antigen retrieval was necessary. It is not clear if the antibody was directly conjugated, or if a secondary antibody was applied.

Confocal imaging and analysis

To prepare tissue for confocal imaging, experimental and control groups of WT and TLR2-deficient mice were sacrificed and perfused in situ with 2% paraformaldehyde. Immediately following the perfusion, the kidneys were dissected, removed, and fixed in a solution of 4% paraformaldehyde overnight. The next morning, the kidneys were placed through a sucrose gradient and then frozen in OCT medium (Sakura). Sections (6 μm thick) were stained with anti-TLR2... (see figure 1).

I found this reference and others by searching via Google Scholar, using the keywords "T2.5" (the clone ID) and TLR2, and formalin or paraformaldehyde.

I hope this helps. Please let me know if you have any questions.

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Question
Answer

Thank you for contacting us. So far, the one reference I have found that used this antibody with frozen tissue sections used acetone as the fixative, not PFA.

Kidneys were snap frozen in liquid nitrogen and stored at –70°C prior to use. For histological examination of TLR2, kidneys were cut into 6-μm sections, fixed for 10 minutes with ice-cold acetone, and air dried. The sections were incubated in 5% normal goat serum (DakoCytomation) for 10 minutes and then exposed overnight to anti-TLR2 mAb T2.5 in PBS.

Leemans, J et al; Renal-associated TLR2 mediates ischemia/reperfusion injury in the kidney. J Clin Invest 2005 115: 2894

I have asked for more data from our source for this clone and will forward this when I receive it. Can you please confirm the species of your samples? I am assuming mouse embryo.

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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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