FSL-1 Synthetic diacylated lipoprotein - TLR2/TLR6 ligand Catalog # tlrl-fsl For research use only Version # 11F09-MM

Product information metHodS content: Preparation of stock solution (100 µg/ml) • 100 µg lipopeptide FSL-1 Stimulation of TLR2/TLR6 can be achieved with 1 ng to 100 ng/ml FSL-1. • 1.5 ml sterile endotoxin-free water - Add 1 ml endotoxin-free water (provided) and vortex until complete Storage and stability: solubilisation. - FSL-1 is provided lyophilized and shipped at room temperature. Store Note: FSL-1 is water soluble in contrast to MALP2. at 4˚C. - Prepare further dilutions by adding the appropriate amount of endotoxin- - Upon resuspension, store at 4°C. Resuspended product is stable for 6 free water. months at 4°C.

deScriPtion O CH3(CH2)14

FSL-1 (Pam2CGDPKHPKSF) is a synthetic lipoprotein that represents the N-terminal part of the 44-kDa lipoprotein LP44 of Mycoplasma O NH2 salivarium1. The framework structure of FSL-1 is the same as that of O

S Gly-Asp-Pro-Lys-His-Pro-Lys-Ser-Phe MALP-2, a Mycoplasma fermentans derived lipopeptide (LP), but they O

CH3(CH2)14 differ in the amino acid sequence and length of the peptide portion2. O Mycoplasmal LP, such as FSL-1 and MALP-2, contain a lipolyated N-terminal diacylated cysteine residue, whereas bacterial LP contain a fSL-1 stimulation triacylated one. This structural difference plays a crucial role in the initial - Transfect your cell line with an NF-kB-inducible reporter plasmid, i.e. a recognition of microbial LP by the host innate immune system. plasmid carrying a reporter gene, such as SEAP or luciferase, under the Mycoplasmal LP, such as FSL-1, are recognized by TLR2 and TLR6, control of an NF-kB-inducible ELAM-1 (E-selectin) promoter. whereas bacterial LP and Pam3CSK4, a synthetic LP, are recognized by Note: InvivoGen provides pNiFty, a family of NF-kB-inducible reporter TLR2 and TLR13. FSL-1 stimulation induces a MyD88-dependent plasmids that can be transfected transiently (pNiFty) or stably (pNiFty2). pNiFty signaling cascade leading to the activation of NF-kB and the production of plasmids are available either with the SEAP or luciferase reporter genes. proinflammatory cytokines. If your cell line does not naturally express TLR2 and TLR6, cotransfect with a plasmid co-expressing both TLRs, such as pDUO-TLR6/TLR2 FSL-1 exhibits a higher stimulatory activity than MALP2 when tested for (see Related Products). its ability to induce the production of TNFa in macrophages and NF-kB in Note: Alternatively, evaluate TLR2 activation using reporter cells, such as TLR2/TLR6 transfected HEK293 cells. InvivoGen’s HEK-Blue™ hTLR2 cells which express the human TLR2 and SEAP reporter genes. NF-kB production in these cells can be easily quantified using 1. Shibata, Ki. et al., 2000. The N-terminal lipopeptide of a 44-kDa membrane-bound a SEAP detection medium, such as QUANTI-Blue™ or HEK-Blue™ Detection. lipoprotein of Mycoplasma salivarium is responsible for the expression of intercellular - Twenty-four to forty-eight hours after transfection, stimulate cells adhesion molecule-1 on the cell surface of normal human gingival fibroblasts. J. Immunol. with 10 ng to 100 ng/ml FSL-1 for 6 hours to 24 hours. 165:6538–6544. 2. okusawa t. et al., 2004. Relationship between Structures and Biological Activities of Mycoplasmal Diacylated Lipopeptides and Their Recognition by - Determine FSL-1 stimulation on TLR2/TLR6 by assessing reporter gene Toll-Like Receptors 2 and 6. Infect Immun. 72(3): 1657-1665. 3. takeuchi o. et al., 2001. expression using the appropriate detection system. Discrimination of bacterial lipoproteins by Toll-like receptor 6. Int Immunol. 13(7):933-40. reLated ProductS cHemicaL ProPertieS chemical name: (S,R)-(2,3-bispalmitoyloxypropyl)-Cys-Gly-Asp- Product catalog code Pro-Lys-His-Pro-Lys-Ser-Phe TM formula: C84H140N14O18S HEK-Blue hTLR2 hkb-htlr2 pNiFty-Luc (AmpR) pnifty-luc molecular weight: 1666.2 pNiFty-SEAP (AmpR) pnifty-seap Solubility: 10 mg/ml in water pNiFty2-Luc (ZeoR) pnifty2-luc pNiFty2-SEAP (ZeoR) pnifty2-seap pDUO-hTLR6/2 (human genes) pduo-htlr6tlr2 pDUO-mTLR6/2 (mouse genes) pduo-mtlr6tlr2

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