FSL-1

Synthetic diacylated lipoprotein - TLR2/TLR6 ligand

Catalog # tlrl-fsl

For research use only

Version # 11F09-MM

Product information metHodS

content: Preparation of sock 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
FSL-1 (Pam2CGDPKHPKSF) is a synthetic lipoprotein that represents the
N-terminal part of the 44-kDa lipoprotein LP44 of Mycoplasma
salivarium1. The framework structure of FSL-1 is the same as that of
MALP-2, a Mycoplasma fermentans derived lipopeptide (LP), but they
differ in the amino acid sequence and length of the peptide portion2.
Mycoplasmal LP, such as FSL-1 and MALP-2, contain a lipolyated
N-terminal diacylated cysteine residue, whereas bacterial LP contain a
triacylated one. This structural difference plays a crucial role in the initial
recognition of microbial LP by the host innate immune system.
Mycoplasmal LP, such as FSL-1, are recognized by TLR2 and TLR6,
whereas bacterial LP and Pam3CSK4, a synthetic LP, are recognized by
TLR2 and TLR13. FSL-1 stimulation induces a MyD88-dependent
signaling cascade leading to the activation of NF-kB and the production of
proinflammatory cytokines.
FSL-1 exhibits a higher stimulatory activity than MALP2 when tested for
its ability to induce the production of TNFa in macrophages and NF-kB in
TLR2/TLR6 transfected HEK293 cells.
1. Shibata, Ki. et al., 2000. The N-terminal lipopeptide of a 44-kDa membrane-bound
lipoprotein of Mycoplasma salivarium is responsible for the expression of intercellular
adhesion molecule-1 on the cell surface of normal human gingival fibroblasts. J. Immunol.
165:6538–6544. 2. okusawa t. et al., 2004. Relationship between Structures and
Biological Activities of Mycoplasmal Diacylated Lipopeptides and Their Recognition by
Toll-Like Receptors 2 and 6. Infect Immun. 72(3): 1657-1665. 3. takeuchi o. et al., 2001.
Discrimination of bacterial lipoproteins by Toll-like receptor 6. Int Immunol. 13(7):933-40.
CH3(CH2)14 O
O
NH2
O
O S Gly-Asp-Pro-Lys-His-Pro-Lys-Ser-Phe
CH3(CH2)14
O
fSL-1 stimulation
- Transfect your cell line with an NF-kB-inducible reporter plasmid, i.e. a
plasmid carrying a reporter gene, such as SEAP or luciferase, under the
control of an NF-kB-inducible ELAM-1 (E-selectin) promoter.
Note: InvivoGen provides pNiFty, a family of NF-kB-inducible reporter
plasmids that can be transfected transiently (pNiFty) or stably (pNiFty2). pNiFty
plasmids are available either with the SEAP or luciferase reporter genes.
If your cell line does not naturally express TLR2 and TLR6, cotransfect
with a plasmid co-expressing both TLRs, such as pDUO-TLR6/TLR2
(see Related Products).
Note: Alternatively, evaluate TLR2 activation using reporter cells, such as
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
a SEAP detection medium, such as QUANTI-Blue™ or HEK-Blue™ Detection.
- Twenty-four to forty-eight hours after transfection, stimulate cells
with 10 ng to 100 ng/ml FSL-1 for 6 hours to 24 hours.
- Determine FSL-1 stimulation on TLR2/TLR6 by assessing reporter gene
expression using the appropriate detection system.

reLated ProductS
cHemicaL ProPertieS
chemical name: (S,R)-(2,3-bispalmitoyloxypropyl)-Cys-Gly-Asp- Product catalog code
Pro-Lys-His-Pro-Lys-Ser-Phe
formula: C84H140N14O18S HEK-Blue hTLR2
TM
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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