Pam3CSK4, TLR1 and TLR2 Ligand

Pam3CSK4, TLR1 and TLR2 Ligand [NBP2-25297] - 293 cells were transfected with pCMV/TLR1-2 plasmid and pNF-kB/SEAP plasmid using Lipofectamin 2000. After 48 hrs of transfection, 50 ng/ml of Pam3CSK4 was added. Cells were incubated at 37C for 24 hrs. Transfected cell supernatant was collected and analyzed using the NF-kB SEAPorter Assay kit. pCMV/SEAP plasmid was used to check transfection efficiency.

Pam3CSK4, TLR1 and TLR2 Ligand [NBP2-25297] - Evaluation of the ligand activity on TLR2/NF-kB SEAPorter HEK 293 cell line. IML-102 cell line is a stably co-transfected cell line that expresses full-length human Toll-like receptor 2 (TLR2) and the secreted alkaline phosphatase reporter gene under the transcriptional control of an NF-KB response element. IML-102 cells were plated in 96-well plates at 5 x 10^4 cells/well. After 16 h, cells were stimulated with various amounts of for 24 h. SEAP was analyzed using the SEAPorter Assay Kit . Data Summary: specifically activated the TLR2/1-depedent NF-KB/SEAP reporter cells in a dose dependent manner (Note: HEK 293 cells endogenously express TLR1).

Pam3CSK4, TLR1 and TLR2 Ligand [NBP2-25297] - Differentiated human macrophages were treated with or without 50 ng/mL Pam3CSK4 for 2 h, and the release of IL-6 in culture supernatants of cells was measured by ELISA kit.

• Reactivity: Hu, Mu, Ba, Ch
• Applications: Func
• Concentration: Please see the protocols for proper use of this product. If no protocol is available, contact technical services for assistance.

Pam3CSK4, TLR1 and TLR2 Ligand Summary

• Description: Pam3CSK4, TLR1 and TLR2 Ligand is a synthetic tripalmitoylated lipohexapeptide analog of the immunologically active N-terminal portion of bacterial lipoprotein. It activates monocytes and macrophages and is also a potent activator of proinflammatory transcription factor NF-kB. Pam3CSK4, TLR1 and TLR2 Ligand is recognized by a heterodimer formed between TLR1 and TLR2.Pam3CSK4, TLR1 and TLR2 Ligand stimulation: If your cell line does not naturally express TLR1+ TLR2, co-transfect with plasmids pCMV/TLR1and pCMV/TLR2. Forty-eight hrs after transfection, stimulate cells with 10 to 100 ng/ml of Pam3CSK4 for 6-24 hrs. Determine Pam3CSK4 stimulation using appropriate detection assays.
• Specificity: Pam3CSK4, TLR1 and TLR2 Ligand

Applications/Dilutions

• Dilutions:
  • Functional
  • In vitro assay
  • In vivo assay
  • Ligand Activation

Reactivity Notes

Human reactivity reported in scientific literature (PMID: 24105263). Mouse reactivity reported in scientific literature (PMID: 25511699) Use in Chicken reported in scientific publication (PMID: 32733155). Use in Bacteria reported in scientific publication (PMID: 31482074).

Packaging, Storage & Formulations

• Storage: Store at 4C short term. Aliquot and store at -20C long term. Avoid freeze-thaw cycles.
• Buffer: Sterile Water
• Concentration: Please see the protocols for proper use of this product. If no protocol is available, contact technical services for assistance.

Alternate Names for Pam3CSK4, TLR1 and TLR2 Ligand

• Pam3CSK4 TLR1 and TLR2 Ligand
• Pam3CSK4
• TLR1 and TLR2 Ligand

Limitations

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Support products are guaranteed for 6 months from date of receipt.

Publications for Pam3CSK4, TLR1 and TLR2 Ligand (NBP2-25297)(30)

Publications using NBP2-25297

• Yamamoto M, Miyoshi M, Morioka K et al. Anti-nucleolin aptamer, iSN04, inhibits the inflammatory responses in C2C12 myoblasts by modulating the ?-catenin/NF-?B signaling pathway Biochemical and biophysical research communications 2023-04-26 [PMID: 37127012]
• Liu Y, Diamond SL. Activation of Most Toll-Like Receptors in Whole Human Blood Attenuates Platelet Deposition on Collagen under Flow Journal of Immunology Research 2023-01-17 [PMID: 36703865]
• Horii T, Orikawa Y, Ohira Y et al. Peptidoglycan-Like Components in Z-100, Extracted from Mycobacterium tuberculosis Strain Aoyama B, Increase IL-12p40 via NOD2 Journal of Immunology Research 2022-06-22 [PMID: 35785036] (LA)
• Kawahara T, Ito A, Kiso A, Kawamoto F Inhibitory effect of strawberry geranium (Saxifraga stolonifera) on Toll-like receptor 2-mediated inflammatory response in human skin keratinocytes Journal of Ethnopharmacology 2021-04-01 [PMID: 33819504]
• Terada T, Nii T, Isobe N, Yoshimura Y Effects of Toll-like Receptor Ligands on the Expression of Proinflammatory Cytokines and Avian beta-defensins in Cultured Chick Intestine J Poult Sci 2020-07-25 [PMID: 32733155] (Chicken)
• Kanoh H, Nitta T, Go S et al. Homeostatic and pathogenic roles of GM3 ganglioside molecular species in TLR4 signaling in obesity EMBO J. 2020-05-07 [PMID: 32378734] (Mouse)
• Effects of TLR Ligands on the Expression of Cytokines and Possible Role of NFkB in its Process in the Theca of Chicken Follicles Kang Y, Nii T, Isobe N J Poult Sci [PMID: 32055188] (Func, Chicken)
• Yamaguchi M, Hirose Y, Takemura M, et al Streptococcus pneumoniae Evades Host Cell Phagocytosis and Limits Host Mortality Through Its Cell Wall Anchoring Protein PfbA. Front Cell Infect Microbiol. 2019-08-20 [PMID: 31482074] (Bacteria)
  
  Details:
  Mice were infected with S. pneumoniae.
• Sun L, Hult EM, Cornell TT et al. Loss of myeloid-specific protein phosphatase 2A enhances lung injury and fibrosis and results in IL-10 dependent sensitization of epithelial cell apoptosis Am. J. Physiol. Lung Cell Mol. Physiol. 2019-03-06 [PMID: 30838865] (Func, Mouse)
• Nitta T, Kanoh H, Inamori KI et al. Globo-series glycosphingolipids enhance Toll-like receptor 4-mediated inflammation and play a pathophysiological role in diabetic nephropathy. Glycobiology. 2018-11-22 [PMID: 30476082] (Mouse)
• Nihashi Y, Ono T, Kagami H, Takaya T. Toll-like receptor ligand-dependent inflammatory responses in chick skeletal muscle myoblasts. J. Orthop. Res. 2018-10-31 [PMID: 30389519]
• Kikuchi J, Kuroda Y, Koyama D et al. Myeloma Cells are Activated in Bone Marrow Microenvironment by the CD180/MD-1 Complex which Senses Lipopolysaccharide Cancer Res. 2018-01-23 [PMID: 29363546] (Mouse)
• Parthasarathy G, Philipp MT. Receptor tyrosine kinases play a significant role in human oligodendrocyte inflammation and cell death associated with the Lyme disease bacterium Borrelia burgdorferi. J Neuroinflammation 2017-05-30 [PMID: 28558791] (Human)
• Wang P, Zeng F, He L et al. Alteration of the immune status of umbilical cord mesenchymal stem cells stimulated by TLR1/2 agonist, Pam3Csk Mol Med Rep 2016-07-13 [PMID: 27431529]
• Yanai S, Tokuhara D, Tachibana D et al. Diabetic pregnancy activates the innate immune response through TLR5 or TLR1/2 on neonatal monocyte J. Reprod. Immunol. 2016-06-22 [PMID: 27351455] (In vitro, Human)
• Croasdell A, Sime PJ, Phipps RP Resolvin D2 decreases TLR4 expression to mediate resolution in human monocytes FASEB J. 2016-09-30 [PMID: 27256622] (LA, LA, Human)
• Sigola LB, Fuentes AL, Millis LM et al. Effects of toll-Like receptor ligands on RAW 264.7 macrophage morphology and zymosan phagocytosis. Tissue and Cell. 2016-04-22 [PMID: 27157550] (Func, Mouse)
• Mastorci K, Muraro E, Pasini E et al. Toll-Like Receptor 1/2 and 5 Ligands Enhance the Expression of Cyclin D1 and D3 and Induce Proliferation in Mantle Cell Lymphoma. PLoS ONE. 2016-04-29 [PMID: 27123851] (Func, Human)
  
  Details:
  This citation used the FITC version of this antibody.
• Phongsisay V, Iizasa E, Hara H, Yoshida H. Evidence for TLR4 and FcRgamma-CARD9 activation by cholera toxin B subunit and its direct bindings to TREM2 and LMIR5 receptors. Mol. Immunol. 2015-05-25 [PMID: 26021803] (Func, In vitro, LA, Human, Mouse)
  
  Details:
  Pam3CSK4, TLR1 and TLR2 Ligand was used at 50-100 ng/ml concentration for the stimulation/ligand activation application in experiments involving TLR4 reporter cell, W / MyD88?/? mouse bone marrow-derived macrophages /BMDMs and NF-?B reporter cells /T?BRed cells (reporter cells express a fluorescent marker DsRed under the control of NF-?B in human monocytes THP1). See full text for detailed protocols and results.
• Nohmi K, Tokuhara D, Tachibana D et al. Zymosan Induces Immune Responses Comparable with Those of Adults in Monocytes, Dendritic Cells, and Monocyte-Derived Dendritic Cells from Cord Blood. J. Pediatr. 2015-05-06 [PMID: 25957979] (LA, Func, In vitro, Human)
  
  Details:
  Zymosan, TLR2 ligand (Imgenex IMG-2212) was used for in-vitro stimulation experiments involving human heparinized cord or adult blood Monocytes, peripheral blood dendritic cells (DCs) and monocyte-derived DCs (MoDCs). Zymosan was employed at 1 ug/mL concentration on Monocytes as well as on MoDCs and at 0.5 ug/mL on DCs. See full text for experimental details and results.
• Kropp KA, Hsieh WY, Isern E et al. A Temporal Gate for Viral Enhancers to Co-opt Toll-Like-Receptor Transcriptional Activation Pathways upon Acute Infection PLoS Pathog. 2015-04-01 [PMID: 25856589] (Func)
• Phongsisay V, Hara H, Fujimoto S. Toll-like receptors recognize distinct proteinase-resistant glycoconjugates in Campylobacter jejuni and Escherichia coli. Mol. Immunol. 2015-03-01 [PMID: 25511699] (In-vitro, Func, Mouse)
• Effects of delayed hypothermia on time?dependent microglial production of inflammatory and anti?inflammatory factors. Matsui T, Kida H, Iha T et al. Folia Neuropathol [PMID: 25118900] (Func, Mouse)
• Phongsisay V, Iizasa E, Hara H, Yamasaki S. LMIR5 extracellular domain activates myeloid cells through Toll-like receptor 4 Mol. Immunol. 2014-07-05 [PMID: 25004110] (In vitro)
  
  Details:
  Pam3CSK4, TLR1 and TLR2 Ligand/Pam3 used at 100 ng/ml concentration on bone marrow-derived macrophage/BMDMs wild type (WT) and gene-deficient mice (-/-) in experiments involving stimulation of cells with 1.2 g/ml fusion protein, 50 g/ml zymosan, or 10 ng/ml LPS for 24 h followed by detection of cytokines production with ELISA (Fig. 3)
• Alpy F, Rousseau A, Schwab Y et al. Cystic fibrosis CFBE41o- cells contain TLR1 SNP I602S and fail to respond to Mycobacterium abscessus. J Cell Sci 2013-12-01 [PMID: 24105263] (In vitro, Human)
• Kusagaya H, Fujisawa T, Yamanaka K et al. Toll-like receptor-mediated airway IL-17C enhances epithelial host defense in an autocrine/paracrine manner. Am. J. Respir. Cell Mol. Biol. 2014-01-01 [PMID: 23944933] (In vitro, Human)
  
  Details:
  TLR ligand treatment (primary normal human bronchial epithelial cells), Figs 1, S1. CpG-ODN was used at 10 ug/ml.
• Kempaiah P, Davidson LB, Perkins DJ, Byrd TF. Cystic fibrosis CFBE41o- cells contain TLR1 SNP I602S and fail to respond to Mycobacterium abscessus. J Cyst Fibros. 2013-02-08 [PMID: 23403223]
• Yokoyama R, Itoh S, Kamoshida G et al. Staphylococcal superantigen-like protein 3 binds to the Toll-like receptor 2 extracellular domain and inhibits cytokine production induced by Staphylococcus aureus, cell wall component, or lipopeptides in murine macrophages. Infect Immun. 2012-08-01 [PMID: 22665377]
• Furuhashi K, Suda T, Hasegawa H et al. Mouse lung CD103+ and CD11bhigh dendritic cells preferentially induce distinct CD4+ T-cell responses. Am J Respir Cell Mol Biol. 2012-02-01 [PMID: 21908266]
  
  Details:
  PAM3CSk4(IMG-2201) & CpG-ODN (IMG-2209Mpt). CD103+ and CD11bhigh mouse lung dendritic (LDCs) cells were stimulated with LPS and with or without PAM3CSk4 and CpG-ODN. Cytokine levels were measured by ELISA, Fig 7.
• Gillaux C, Mehats C, Vaiman D et al. Functional screening of TLRs in human amniotic epithelial cells. J Immunol. 2011-09-01 [PMID: 21775685]
  
  Details:
  TLR ligands: TLR1/2 (IMG-2201), TLR3 (IMG-2203), TLR4 (IMG-2204), TLR5 (IMG-2205), TLR6/2 (IMG-2206), TLR7 (IMG-2207), TLR9 (IMG-2209Hpt). The effects of ligand stimulation was measured by various readout assays, refer to the figures for details (Figs 2-8, S1).

Review for Pam3CSK4, TLR1 and TLR2 Ligand (NBP2-25297) (1)

Average Rating: 5

(Based on 1 review)

reviewed by: Eric Wang 06/21/2021

Summary

• Lot: 11281706