NK-4 was synthesized at Functional Dyes Unit, Hayashibara Co., Ltd. (Okayama Japan). A stock solution of 10 mM NK-4 was prepared with DMSO and stored frozen at − 80 °C. Endotoxin content in 100 μM NK-4 was below detection limits (0.00625 endotoxin units/ml), as determined according to a protocol listed in the Japanese Pharmacopoeia using an Endospecy ES-50 M set (Seikagaku Co., Tokyo, Japan). Final concentrations of DMSO at 0.05% or less did not affect the results of the experiments. PMA and LPS (E. coli O55:B5) were purchased from Sigma-Aldrich Japan (Tokyo, Japan). Pam3CSK4 hydrochloride was purchased from InvivoGen (San Diego, CA). Poly(I:C) was purchased from Calbiochem (La Jolla, CA). Recombinant human IL-4 and IL-13 were purchased from R&D Systems (Minneapolis, MN). Recombinant annexin V was purchased from BD Biosciences (Franklin Lakes, NJ). Wortmannin was purchased from Wako Pure Chemical (Osaka, Japan). Human TNF-α, and monoclonal antibodies (mAb) for the human TNF-α ELISA were prepared and purified in our laboratories.
Pam3csk4 hydrochloride
Manufactured by InvivoGen
Pam3CSK4 hydrochloride is a synthetic triacylated lipoprotein that acts as a Toll-like receptor 2 (TLR2) agonist. It is commonly used in research to stimulate TLR2-mediated immune responses.
Automatically generated - may contain errors
2 protocols using pam3csk4 hydrochloride
1
Synthesis and Validation of NK-4 Compound
2
Endotoxin-free NK-4 Compound Synthesis
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Reagents NK-4 was synthesized at Functional Dyes Unit, Hayashibara Co., Ltd. (Okayama Japan). A stock solution of 10 mM NK-4 was prepared with DMSO and stored frozen at -80°C. Endotoxin content in 100 mM NK-4 was below detection limits (0.00625 endotoxin units/ml), as determined according to a protocol listed in the Japanese Pharmacopoeia using an Endospecy ES-50M set (Seikagaku Co., Tokyo, Japan). Final concentrations of DMSO at 0.05% or less did not affect the results of the experiments. PMA and LPS (E. coli O55:B5) were purchased from Sigma-Aldrich Japan (Tokyo, Japan). Pam3CSK4 hydrochloride was purchased from InvivoGen (San Diego, CA). Poly(I:C) was purchased from Calbiochem (La Jolla, CA). Recombinant human IL-4 and IL-13 were purchased from R&D Systems (Minneapolis, MN). Recombinant annexin V was purchased from BD Biosciences (Franklin Lakes, NJ). Wortmannin was purchased from Wako Pure Chemical (Osaka, Japan). Human TNF-a, and monoclonal antibodies (mAb) for the human TNF-a ELISA were prepared and puri ed in our laboratories.
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