| Size | Price | |
|---|---|---|
| 500mg | ||
| 1g | ||
| Other Sizes |
Pam3CSK4-Biotin is a biotinylated Pam3CSK4 acting as a novel and potent Toll-like receptor 1/2 (TLR1/2) agonist.
| Targets |
- Toll-like receptor 2 (TLR2)-TLR1 heterodimer
- No IC50/Ki/EC50 values for Pam3CSK4-Biotin were reported in the study. The parent compound Pam3CSK4 is a TLR2-TLR1 agonist with an EC50 of 43.2 ± 5.6 nM for equine TLR2-TLR1 activation [1]
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|---|---|
| ln Vitro |
- TLR2-TLR1 signaling activation:
1. NF-κB reporter gene activation: HEK-Blue™ cells expressing human TLR2-TLR1 were treated with Pam3CSK4 (100 ng/mL) or LTA (1 μg/mL) for 24 hours. Both ligands induced NF-κB-dependent SEAP (secreted embryonic alkaline phosphatase) activity, which was abolished by clathrin/dynamin inhibitors (chlorpromazine, dynasore) [1] 2. Cytokine induction: Human primary monocytes treated with Pam3CSK4 (100 ng/mL) for 24 hours showed increased TNF-α secretion (ELISA), which was blocked by endocytosis inhibitors [1] |
| Enzyme Assay |
- NF-κB reporter gene assay for TLR2-TLR1 activation [1]:
1. Cell culture: HEK-Blue™ cells (expressing TLR2-TLR1 and SEAP reporter) were seeded in 96-well plates (1×10⁵ cells/well). 2. Drug treatment: Cells were incubated with Pam3CSK4 (10–1000 ng/mL) or LTA (0.1–10 μg/mL) for 24 hours at 37°C, 5% CO₂. 3. Activity detection: SEAP activity in cell supernatants was measured using QUANTI-Blue™ reagent, with absorbance read at 655 nm. Relative activity was normalized to vehicle control. |
| Cell Assay |
- Monocyte endocytosis and cytokine induction assay [1]:
1. Cell isolation: Human peripheral blood monocytes were isolated via CD14⁺ magnetic beads and cultured in RPMI 1640 with 10% FBS. 2. Drug treatment: Monocytes were pre-treated with endocytosis inhibitors (chlorpromazine, dynasore, chloroquine) for 45 minutes, then stimulated with Pam3CSK4 (100 ng/mL) or LTA (1 μg/mL) for 4–24 hours. 3. Analysis: TNF-α mRNA expression (qPCR) and protein secretion (ELISA) were measured. Internalization of Pam3CSK4 was visualized via confocal microscopy using Alexa Fluor-labeled Pam3CSK4 (not biotinylated) [1] |
| References | |
| Additional Infomation |
Key findings: This study shows that TLR2 ligands (Pam3CSK4 and LTA) can induce NF-κB activation in the endosome compartment of human monocytes. This process requires clathrin/dynein-dependent endocytosis and CD14-mediated internalization [1]
- No biotinylated analogues used: This study did not use Pam3CSK4-biotin, but focused on unmodified Pam3CSK4. Biotinylation of Pam3CSK4 is a common modification used to track ligand-receptor interactions (e.g., streptavidin-based detection), but its specific role was not investigated in this study [3,6]. - Mechanism analysis: Endosomal localization of TLR2-TLR1 is crucial for signal transduction because cell surface TLR2 itself cannot activate NF-κB without internalization [1]. - Species relevance: The results are applicable only to human monocytes expressing human TLR2-TLR1 and HEK-Blue™ cells [1]. |
| Molecular Formula |
C103H192N14O17S2
|
|---|---|
| Molecular Weight |
1962.85
|
| Exact Mass |
1961.40313
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| CAS # |
1429504-10-8
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| PubChem CID |
90488959
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| SequenceShortening |
XSKKKX
|
| Appearance |
Typically exists as solid at room temperature
|
| LogP |
17.5
|
| Hydrogen Bond Donor Count |
16
|
| Hydrogen Bond Acceptor Count |
22
|
| Rotatable Bond Count |
98
|
| Heavy Atom Count |
98
|
| Complexity |
3160
|
| Defined Atom Stereocenter Count |
9
|
| SMILES |
C([C@@H]1SC[C@]2([H])NC(N[C@@]21[H])=O)CCCC(=O)NCCCCCC(=O)NCCCCCC(=O)NCCCC[C@@H](C(=O)O)NC(=O)[C@H](CCCCN)NC(=O)[C@H](CCCCN)NC(=O)[C@H](CCCCN)NC(=O)[C@H](CO)NC(=O)[C@@H](NC(=O)CCCCCCCCCCCCCCC)CSCC(COC(=O)CCCCCCCCCCCCCCC)OC(=O)CCCCCCCCCCCCCCC
|
| InChi Key |
GVXJPWKMFMNHRE-XHHKIZCASA-N
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| InChi Code |
InChI=1S/C103H192N14O17S2/c1-4-7-10-13-16-19-22-25-28-31-34-37-42-67-93(122)110-88(79-135-78-81(134-95(124)69-46-39-36-33-30-27-24-21-18-15-12-9-6-3)77-133-94(123)68-45-38-35-32-29-26-23-20-17-14-11-8-5-2)101(129)115-86(76-118)100(128)113-83(60-50-54-71-105)98(126)111-82(59-49-53-70-104)97(125)112-84(61-51-55-72-106)99(127)114-85(102(130)131)62-52-58-75-109-91(120)65-44-41-56-73-107-90(119)64-43-40-57-74-108-92(121)66-48-47-63-89-96-87(80-136-89)116-103(132)117-96/h81-89,96,118H,4-80,104-106H2,1-3H3,(H,107,119)(H,108,121)(H,109,120)(H,110,122)(H,111,126)(H,112,125)(H,113,128)(H,114,127)(H,115,129)(H,130,131)(H2,116,117,132)/t81?,82-,83-,84-,85-,86-,87-,88-,89-,96-/m0/s1
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| Chemical Name |
(2S)-6-[6-[6-[5-[(3aS,4S,6aR)-2-oxo-1,3,3a,4,6,6a-hexahydrothieno[3,4-d]imidazol-4-yl]pentanoylamino]hexanoylamino]hexanoylamino]-2-[[(2S)-6-amino-2-[[(2S)-6-amino-2-[[(2S)-6-amino-2-[[(2S)-2-[[(2R)-3-[2,3-di(hexadecanoyloxy)propylsulfanyl]-2-(hexadecanoylamino)propanoyl]amino]-3-hydroxypropanoyl]amino]hexanoyl]amino]hexanoyl]amino]hexanoyl]amino]hexanoic acid
|
| Synonyms |
Pam3CSK4 Biotin; 1429504-10-8; orb1984076; AKOS024458267; PD079725; (2S,3Z,5S,6Z,8S,9Z,11S,12Z,14S,15Z,17R)-5,8,11-tris(4-aminobutyl)-4,7,10,13,16-pentahydroxy-2-(4-((Z)-(1-hydroxy-6-((Z)-(1-hydroxy-6-((Z)-(1-hydroxy-5-((3aR,6S,6aS)-2-hydroxy-3a,4,6,6a-tetrahydro-1H-thieno[3,4-d]imidazol-6-yl)pentylidene)amino)hexyli
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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 | 0.5095 mL | 2.5473 mL | 5.0946 mL | |
| 5 mM | 0.1019 mL | 0.5095 mL | 1.0189 mL | |
| 10 mM | 0.0509 mL | 0.2547 mL | 0.5095 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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