TIRAP (TLR2 and TLR4) Inhibitor Peptide Set

Human, Mouse

Flow Cytometry, Functional Assay

TLR2 and TLR4 Inhibitor Peptide: COBRA
TLR2 and TLR4 Inhibitor interferes with interaction between TIRAP/Mal and TIR domain of TLR2 or TLR4.

Flow Cytometry (reported in scientific literature (PMID 27838489))
In vitro assay (reported in scientific literature (PMID 26908090))

The inhibitor is used in assays to inhibit TLR2 and TLR4 signaling. We recommend an initial titration of the inhibitor from 0-50 uM for in vitro assays along with control of which concentrations should be mirror inhibitor concentrations. Inhibitor and control should be preincubated with cells prior to ligand activation to allow sufficient time for the peptides to enter from the media into the cell. We typically preincubate with inhibitor and control for 1 h prior to TLR2 or TLR4 activation (Figures 1 and 2); however, optimal preincubation times may vary between model systems. The TLR2/NF-kB/SEAPorter™ cell line and TLR4/MD2/CD14/NF-kB/SEAPorter™ cell line are a useful positive control model system for studying inhibition of TLR2 and TLR4 activation, respectively (Figures 1 through 3).

Preparation of 2.5 mM peptide stock solutions
TLR2 / TLR4 Inhibitor Peptide: A final volume of 100 ul will make a 2.5 mM stock solution. Add 100 ul sterile water to the tube of peptide. Carefully pipet to ensure all of the peptide is dissolved and briefly spin the tube before opening.

Antennapedia Control Peptide: A final volume of 170 ul will make a 2.5 mM stock solution. Add 170 ul sterile water to the tube of peptide. Carefully pipet to ensure all of the peptide is dissolved and briefly spin the tube before opening.

The stock solutions may be diluted further to make working solutions. Dilute according to the needs for your assay. For example, dilute 2.5 mM stock solutions 1:10 in sterile 1X PBS or cell culture media to make 250 uM working solutions. Working solution should be made fresh daily and not be stored.

Form: White solid
Solubilize the peptides prior to use by preparing 2.5 mM stock solution in sterile water.

Concentration is not relevant for this product. Please see the protocols for proper use of this product.

The product is shipped with polar packs. Upon receipt, store it immediately at the temperature recommended below.

Aliquot and store at -20C or -80C. Avoid freeze-thaw cycles.