| Size | Price | Stock | Qty |
|---|---|---|---|
| 1mg |
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| Other Sizes |
| Targets |
Pam2CSK-4 targets the Toll-like receptor 2 (TLR2) in complex with TLR6. It is a TLR6-independent TLR2 ligand, binding to the TLR2/TLR6 heterodimer. Upon binding, it triggers the MyD88-dependent signaling cascade, leading to the activation of NF-kappaB and MAP kinase pathways. This ultimately results in the production of pro-inflammatory cytokines (e.g., TNF-alpha, IL-6), chemokines, and the expression of co-stimulatory molecules.
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| ln Vitro |
Pam2CSK-4 is a potent agonist with an EC50 of 0.015 ng/mL for activating the TLR2/TLR6 pathway. It induces TNF-alpha production in human mononuclear cells, promotes the proliferation and activation of mouse splenic B cells, and induces the expression of inducible nitric oxide synthase (iNOS) and nitric oxide (NO) in macrophage cell lines (e.g., RAW 264.7) through TBK1 and MyD88. It also activates platelets, promoting aggregation and adhesion.
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| ln Vivo |
Pam2CSK-4 is used in vivo to induce systemic or localized inflammatory responses. For example, intraperitoneal injection of Pam2CSK-4 in mice leads to the production of systemic pro-inflammatory cytokines and can be used to study sepsis-like responses. It is also used in models of skin inflammation, arthritis, and lung injury to understand the role of TLR2 signaling in disease pathology. A typical dose in mice is 10-100 microg per mouse via intraperitoneal injection.
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| Enzyme Assay |
A biochemical binding assay for Pam2CSK-4 measures its ability to activate TLR2 signaling in a reporter cell line, not a direct protein-protein binding. HEK293 cells are stably transfected with human TLR2, TLR6, and an NF-kappaB-driven luciferase reporter gene. Varying concentrations of Pam2CSK-4 are added to the cell culture. After several hours of incubation, the cells are lysed, and the luciferase activity is measured. The EC50 of the agonist is determined as the concentration that gives a half-maximal induction of the reporter signal.
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| Cell Assay |
A cellular activation assay is performed using RAW 264.7 murine macrophages or primary human peripheral blood mononuclear cells (PBMCs). Cells are seeded in a 96-well plate and treated with varying concentrations of Pam2CSK-4 (e.g., 1-1000 ng/mL) for 4-24 hours. After treatment, culture supernatants are collected, and the levels of pro-inflammatory cytokines (e.g., TNF-alpha, IL-6, IL-1beta) are measured by ELISA. For pathway analysis, cells are harvested, lysed, and analyzed by Western blot for the phosphorylation of signaling molecules like p38 MAPK, JNK, and IkappaBalpha.
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| Animal Protocol |
To study TLR2-driven inflammation in vivo, female C57BL/6 mice are injected intraperitoneally with Pam2CSK-4 (e.g., 50-100 ug per mouse in 200 uL of sterile PBS). After 1.5-2 hours, the mice are sacrificed. Blood is collected via cardiac puncture, and serum is isolated to measure systemic levels of cytokines like TNF-alpha and IL-6 by ELISA. Bronchoalveolar lavage (BAL) fluid can be collected for cell differentials to assess lung inflammation. As a positive control for TLR4 activation, LPS is often used.
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| ADME/Pharmacokinetics |
Being a synthetic lipopeptide, Pam2CSK-4 is not expected to have a prolonged half-life or oral bioavailability. It is used via systemic injection (e.g., i.p., i.v., or s.c.) for in vivo experiments. Its activity is localized and acute, as it mimics a pathogen-associated molecular pattern (PAMP) and is rapidly cleared. Specific PK parameters are not studied for this agonist, as it is not a drug candidate.
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| Toxicity/Toxicokinetics |
As a potent TLR2 agonist, Pam2CSK-4 is inherently toxic at high doses, as it induces a strong, acute inflammatory response (septic shock). The primary toxicity is due to the overproduction of pro-inflammatory cytokines (“cytokine storm”). Therefore, its use is strictly for research and is controlled, with doses carefully optimized. It is not for human use. The LD50 in mice would be a measure of its acute toxicity.
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| Additional Infomation |
Pam2CSK-4 is a pure research-grade chemical and is not a therapeutic drug. It is one of the most specific and widely used agonists for studying TLR2-mediated immune responses. It is an invaluable tool for differentiating TLR2 activation from other pattern recognition receptors (like TLR4, whose ligand is LPS) and for probing the molecular mechanisms of innate immunity. As such, it is not approved for clinical use and is exclusively for laboratory research.
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| Molecular Formula |
C65H126N10O12S
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|---|---|
| Molecular Weight |
1271.82075738907
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| Exact Mass |
1270.927
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| CAS # |
868247-72-7
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| PubChem CID |
71457280
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| Appearance |
Typically exists as solid at room temperature
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| Density |
1.1±0.1 g/cm3
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| Boiling Point |
1302.9±65.0 °C at 760 mmHg
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| Flash Point |
741.7±34.3 °C
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| Vapour Pressure |
0.0±0.6 mmHg at 25°C
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| Index of Refraction |
1.518
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| LogP |
12.33
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| Hydrogen Bond Donor Count |
12
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| Hydrogen Bond Acceptor Count |
18
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| Rotatable Bond Count |
65
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| Heavy Atom Count |
88
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| Complexity |
1810
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| Defined Atom Stereocenter Count |
7
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| SMILES |
[C@H](CCCCN)(C(=O)N[C@@H](CCCCN)C(=O)N[C@H](C(=O)O)CCCCN)NC(=O)[C@H](CCCCN)NC(=O)[C@H](CO)NC(=O)[C@@H](N)CSC[C@H](OC(=O)CCCCCCCCCCCCCCC)COC(=O)CCCCCCCCCCCCCCC
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| InChi Key |
LJUIOEFZFQRWJG-GHYFRYPYSA-N
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| InChi Code |
InChI=1S/C65H126N10O12S/c1-3-5-7-9-11-13-15-17-19-21-23-25-27-41-58(77)86-48-51(87-59(78)42-28-26-24-22-20-18-16-14-12-10-8-6-4-2)49-88-50-52(70)60(79)75-57(47-76)64(83)73-54(38-30-34-44-67)62(81)71-53(37-29-33-43-66)61(80)72-55(39-31-35-45-68)63(82)74-56(65(84)85)40-32-36-46-69/h51-57,76H,3-50,66-70H2,1-2H3,(H,71,81)(H,72,80)(H,73,83)(H,74,82)(H,75,79)(H,84,85)/t51-,52+,53+,54+,55+,56+,57+/m1/s1
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| Chemical Name |
(2S)-6-amino-2-[[(2S)-6-amino-2-[[(2S)-6-amino-2-[[(2S)-6-amino-2-[[(2S)-2-[[(2R)-2-amino-3-[(2R)-2,3-di(hexadecanoyloxy)propyl]sulfanylpropanoyl]amino]-3-hydroxypropanoyl]amino]hexanoyl]amino]hexanoyl]amino]hexanoyl]amino]hexanoic acid
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| HS Tariff Code |
2934.99.9001
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| Storage |
Powder -20°C 3 years 4°C 2 years In solvent -80°C 6 months -20°C 1 month |
| Shipping Condition |
Room temperature (This product is stable at ambient temperature for a few days during ordinary shipping and time spent in Customs)
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| Solubility (In Vitro) |
May dissolve in DMSO (in most cases), if not, try other solvents such as H2O, Ethanol, or DMF with a minute amount of products to avoid loss of samples
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| Solubility (In Vivo) |
Note: Listed below are some common formulations that may be used to formulate products with low water solubility (e.g. < 1 mg/mL), you may test these formulations using a minute amount of products to avoid loss of samples.
Injection Formulations
Injection Formulation 1: DMSO : Tween 80： Saline = 10 : 5 : 85 (i.e. 100 μL DMSO stock solution → 50 μL Tween 80 → 850 μL Saline)(e.g. IP/IV/IM/SC) *Preparation of saline: Dissolve 0.9 g of sodium chloride in 100 mL ddH ₂ O to obtain a clear solution. Injection Formulation 2: DMSO : PEG300 ：Tween 80 : Saline = 10 : 40 : 5 : 45 (i.e. 100 μL DMSO → 400 μLPEG300 → 50 μL Tween 80 → 450 μL Saline) Injection Formulation 3: DMSO : Corn oil = 10 : 90 (i.e. 100 μL DMSO → 900 μL Corn oil) Example: Take the Injection Formulation 3 (DMSO : Corn oil = 10 : 90) as an example, if 1 mL of 2.5 mg/mL working solution is to be prepared, you can take 100 μL 25 mg/mL DMSO stock solution and add to 900 μL corn oil, mix well to obtain a clear or suspension solution (2.5 mg/mL, ready for use in animals). View More
Injection Formulation 4: DMSO : 20% SBE-β-CD in saline = 10 : 90 [i.e. 100 μL DMSO → 900 μL (20% SBE-β-CD in saline)] Oral Formulations
Oral Formulation 1: Suspend in 0.5% CMC Na (carboxymethylcellulose sodium) Oral Formulation 2: Suspend in 0.5% Carboxymethyl cellulose Example: Take the Oral Formulation 1 (Suspend in 0.5% CMC Na) as an example, if 100 mL of 2.5 mg/mL working solution is to be prepared, you can first prepare 0.5% CMC Na solution by measuring 0.5 g CMC Na and dissolve it in 100 mL ddH2O to obtain a clear solution; then add 250 mg of the product to 100 mL 0.5% CMC Na solution, to make the suspension solution (2.5 mg/mL, ready for use in animals). View More
Oral Formulation 3: Dissolved in PEG400 (Please use freshly prepared in vivo formulations for optimal results.) |
| Preparing Stock Solutions | 1 mg | 5 mg | 10 mg | |
| 1 mM | 0.7863 mL | 3.9314 mL | 7.8627 mL | |
| 5 mM | 0.1573 mL | 0.7863 mL | 1.5725 mL | |
| 10 mM | 0.0786 mL | 0.3931 mL | 0.7863 mL |
*Note: Please select an appropriate solvent for the preparation of stock solution based on your experiment needs. For most products, DMSO can be used for preparing stock solutions (e.g. 5 mM, 10 mM, or 20 mM concentration); some products with high aqueous solubility may be dissolved in water directly. Solubility information is available at the above Solubility Data section. Once the stock solution is prepared, aliquot it to routine usage volumes and store at -20°C or -80°C. Avoid repeated freeze and thaw cycles.
Calculation results
Working concentration： mg/mL；
Method for preparing DMSO stock solution： mg drug pre-dissolved in μL DMSO (stock solution concentration mg/mL). Please contact us first if the concentration exceeds the DMSO solubility of the batch of drug.
Method for preparing in vivo formulation:：Take μL DMSO stock solution, next add μL PEG300, mix and clarify, next addμL Tween 80, mix and clarify, next add μL ddH2O，mix and clarify.
(1) Please be sure that the solution is clear before the addition of next solvent. Dissolution methods like vortex, ultrasound or warming and heat may be used to aid dissolving.
(2) Be sure to add the solvent(s) in order.
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