Antibodies to Toll-Like Receptor (TLR) Family - FocusOn 004

The innate or natural immune system recognizes a wide spectrum of pathogens without a need for prior exposure. The main cells responsible for innate immunity, monocytes/macrophages and neutrophils, phagocytose microbial pathogens and trigger the innate, inflammatory and specific immune responses. Members of the Toll-like receptor (TLR) family, first discovered in drosophila, have been shown to be pattern recognition receptors, each member recognizing and responding to different microbial components to limit/eradicate invading microbes. Binding of pathogen-associated molecular patterns (PAMP) to TLRs induces the production of reactive oxygen and nitrogen intermediates, initiation of the pro-inflammatory cytokine network and upregulation of costimulatory molecules linking the rapid innate response to the adaptive immunity.

Fig. 1: CD281/TLR1 antibody Cat.-No. AP07365PU-N staining of formalin-fixed paraffin-embedded human small intestine
Fig.2: Immunohistochemical analysis of TLR4 in formalin-fixed paraffin-embedded human testis tissue using TLR4 antibody SM7124P at 5 µg/ml.

The TLR family has significant homology in its cytoplasmic domain to the IL-1 receptor type I. Studies so far show that signaling pathways via TLRs originate from the conserved Toll/IL-1 receptor (TIR) domain. The TIR domain-containing MyD88 acts as a common adapter that induces secretion of inflammatory cytokines such as IL-1, IL-6, IL-8, IL-12 and TNF-alpha, and induction of co-stimulatory molecules. A MyD88-independent pathway also has been reported to induce type I Interferons (IFNs) in the TLR4 and TLR3 signaling pathway. Another TIR domain-containing adapter, TIRAP/Mal has recently been shown to mediate the MyD88-dependent activation in the TLR4 and TLR2 signaling pathway. Thus, individual TLRs may utilize different signaling systems that characterize their specific activities. In addition to induction of the cytokine network, MyD88 binds FADD and triggers apoptosis through the caspase cascade. Hence, activation of the apoptosis pathway via TLRs appears to contribute to the repertoire of defense mechanisms utilized by the innate immune response.

Fig.3: Western blot: CD281/TLR1 antibody Cat.-No. AP09159PU-N staining of mouse spleen tissue whole cell lysate. This IgG fraction of TLR1 antibody shows detection of a predominant band at ~90 kDa corresponding to TLR1 (arrowhead). The predicted MW of TLR1 is 90 kDa. TLR1 was detected using 1.0 µg/ml (Lane A, 2.0 µg/ml (Lane B) and 4.0 µg/ml (Lane C) concentrations of primary AB
Fig.4: Western blot analysis of TLR5 in (A) Ramos, (B) raw, (C) mouse spleen, (D) mouse lung and (E) rat lung whole cell lysate using Cat.-No. SP7197P at 2 µg/ml.
Fig. 5: Western blot analysis with TLR9 antibody Cat.-No. SM7105P in human PBMC (Lane A), human intestine (Lane B), mouse intestine (Lane C) and rat intestine tissue lysates (Lane D) using the AB at a dilution of 3 µg/ml. Lane E shows antibody tested at 5 µg/ml in human MCF7 cell lysate

The ligands for TLR2, TLR3, TLR4, TLR7 and TLR9 have been reported. TLR2, in association with TLR1 and/or TLR6, recognizes and signals bacterial lipoproteins (BLP) and peptidoglycans from Gram-positive bacteria. Recently, it was discovered that TLR3 recognizes dsRNA. LPS from Gram-negative bacteria signals through TLR4. The immune response modifiers, imiquimod and resiquimod, have been reported as agonists for TLR7. Cellular response to the CpG motif of bacterial DNA is mediated by TLR9. The cDNA for human TLR10 codes for a protein containing 811 amino acids. Human TLR10 is closely related to human TLR1 and TLR6. TLR10 is expressed in tissues and cells of the immune system. TLR10, originally identified from a spleen cDNA library, is on chromosome 4p14 and has been found to be expressed in lung and in B-lymphocytes. TLR10 is thought to be a potential asthma candidate gene because early life innate immune responses to ubiquitous inhaled allergens and PAMPs may influence asthma susceptibility.

In addition to the innate immune response, evidence implicates the involvement of the TLR family in a spectrum of systemic disorders following bacterial infections including sepsis, cardiac ischemia, peridontitis, and cerebral palsy. The rapid pace of research in the TLR family will elucidate their functions and potential therapeutic interventions.

 

Acris Antibodies offers a wide range of antibodies for characterizing  members of the TLR family. A short overview is given in the product list, please note that a majority of these antibodies are also available in Biotin- or fluorescence-dye conjugated formats suitable for particular applications.



Key references

 
Akira, S. et al. (2001) Toll-like receptors: critical proteins linking innate and acquired immunity.
Nat Immunol. 8:675-80.
 
Hopkins PA, Sriskandan S.: Mammalian Toll-like receptors: to immunity and beyond.
Clin Exp Immunol. Jun 2005;140(3):395-407.

Further readings

See the PDF-version: FocusOn 004

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Primary Antibodies

Catalog No. Host Iso. Clone Pres. React. Applications  
AM26265PU-N

CD282 / TLR2 antibody

CD282 / TLR2 Mouse IgG2a TL2.3 Purified Can, Hu C, E, F, FN, ICC/IF, WB
0.1 mg / €300.00
  Acris Antibodies GmbH
SM1825BS

CD282 / TLR2 antibody

TLR2 staining in human spleen by using mAb TL2.1 (10µg/ml). Mouse IgG2a TL2.1 Biotin Can, Hu, Mky C, F, ICC/IF, P, WB
50 µg / €240.00
  Acris Antibodies GmbH
AM26252PU-N

CD283 / TLR3 antibody

CD283 / TLR3 Mouse IgG1 TLR3.7 Purified Can, Hu, Ms C, F, FN, ICC/IF, IP, P, WB
0.1 mg / €300.00
  Acris Antibodies GmbH
SM7101P

CD283 / TLR3 antibody

Figure 1. Analysis of TLR3 in (A) Human gut lumen (longitudinal section, transverse region) using SM7101P at 10 µg/ml (Data courtesy Dr. Elizabeth Furrie, University of Dundee) and Mouse spleen tissue using Isotype Control (B) and SM7101P (C) at 5 mg/ml. 
Mouse IgG1 40C1285.6 Purified Can, Hu, Ms C, F, ICC/IF, IP, P, WB
0.1 mg / €360.00
  Acris Antibodies GmbH
SM2043P

CD289 / TLR9 antibody

CD289 / TLR9 Mouse IgG2a 5G5 Purified Can, Hu, Ms C, F, WB
0.25 mg / €480.00
  Acris Antibodies GmbH
SM2043PS

CD289 / TLR9 antibody

CD289 / TLR9 Mouse IgG2a 5G5 Purified Can, Hu, Ms C, F, WB
0.1 mg / €290.00
  Acris Antibodies GmbH
SM2043PT

CD289 / TLR9 antibody

CD289 / TLR9 Mouse IgG2a 5G5 Purified Can, Hu, Ms C, F, WB
25 µg / €190.00
  Acris Antibodies GmbH
SM7105P

CD289 / TLR9 (268-284) antibody





Intracellular flow analysis of TLR9 in human PBMC using 0.5 µg of TLR9 antibody (red) and isotype control antibody (green). (Cat.-No.: SM7105P)



Mouse IgG1 26C593.2 Purified Can, Eq, Hu, Mky, Ms, Rt C, EMSA, F, ICC/IF, P, WB
0.1 mg / €360.00
  Acris Antibodies GmbH
SM7102P

TLR5 antibody

Figure 1. Western blot analysis of TLR5 in A) Ramos and B) Raw cell lysate using SM7102P at 2 ug/ml. 
Mouse IgG2a 85B152.5 Purified Can, Hu, Ms F, WB
0.1 mg / €330.00
  Acris Antibodies GmbH
SM7103P

TLR5 antibody

Western blot analysis of TLR5 in A) Ramos and B) Raw cell lysate using SM7103P at 2 µg/ml. 
Mouse IgG2a 19D759.2 Purified Can, Hu, Ms C, F, P, WB
0.1 mg / €330.00
  Acris Antibodies GmbH

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