| Size | Price | Stock | Qty |
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| 1mg |
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| 5mg |
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| 10mg |
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| Other Sizes |
| Targets |
MLCK[1].
Myosin light chain kinase (MLCK). This peptide is a specific activator of MLCK. Unlike calmodulin (CaM), which requires Ca2+, this peptide activates MLCK in a Ca2+-independent manner. It binds to MLCK and enhances the phosphorylation of the myosin regulatory light chain (LC2), a critical step in the initiation of smooth muscle contraction and cytoskeletal reorganization. |
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| ln Vitro |
In the lack of exogenous calmodulin (CaM), 16-38-thymosin β4 cattle TFA (50 pM) greatly enhances the percent phosphorylation of LC2.
In vitro, 16-38-Thymosin beta4 (cattle) TFA (50 pM) greatly enhances the percentage of phosphorylation of LC2 (myosin light chain 2) in the absence of exogenous calmodulin (CaM). This demonstrates its potent, Ca2+-independent activation of MLCK. It is used to study the non-canonical activation pathways of myosin II. |
| ln Vivo |
The peptide is used in vivo to study vascular and smooth muscle function. When applied locally or administered systemically, it can induce vasoconstriction and modulate gut motility by activating MLCK independent of the traditional Ca2+/CaM pathway. This helps researchers dissect the distinct roles of Ca2+-dependent and Ca2+-independent MLCK activation in physiology.
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| Enzyme Assay |
A non-cell kinase activity assay is performed. Purified MLCK is incubated with the peptide (0.1-1000 nM) and its substrate, the myosin regulatory light chain (LC2), in the presence of ATP. The reaction is carried out in a buffer with chelators (EGTA) to exclude Ca2+. The level of phosphorylated LC2 is measured by Western blot using a phospho-specific antibody (p-MLC2) to determine activation (EC50).
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| Cell Assay |
For cellular assays, smooth muscle cells are seeded in 6-well plates and serum-starved. The cells are then treated with 16-38-Thymosin beta4 (cattle) TFA (1-100 pM) for 5-30 minutes. Cell lysates are collected and analyzed by Western blot for phosphorylation of MLC2 (p-MLC2) and other downstream targets. The peptide induces robust MLC2 phosphorylation independent of Ca2+ influx.
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| Animal Protocol |
For in vivo studies, 16-38-Thymosin beta4 (cattle) TFA can be administered by intravenous (IV) injection to anesthetized rats at doses of 1-50 ug/kg. Blood pressure is monitored via a carotid artery catheter. The peptide is injected as a bolus to observe its vasoactive effects (e.g., mean arterial pressure). Mesenteric arteries can also be mounted in a myograph to measure isometric tension.
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| ADME/Pharmacokinetics |
The TFA salt form is highly soluble in water and DMSO. It has a molecular weight of 2841.10. For in vitro studies, it is dissolved in sterile water or PBS. For in vivo administration, it can be formulated in saline. The peptide has a short half-life (minutes) due to proteolytic degradation and is typically used for acute experiments. Storage: Lyophilized powder at -20degC.
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| Toxicity/Toxicokinetics |
The peptide is generally non-toxic at the concentrations used for research (pM to nM). At high systemic doses, it may cause prolonged vasoconstriction leading to ischemia. The TFA counterion is non-toxic at low concentrations. No formal toxicology studies have been published for this research peptide. It is for research use only.
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| References | |
| Additional Infomation |
16-38-Thymosin beta4 (cattle) TFA is a research-grade peptide tool for studying MLCK activation and smooth muscle contraction. It is not a drug and has no FDA approval for human therapy. It is widely used in vascular biology research to dissect Ca2+-dependent vs. Ca2+-independent pathways. Key references describe its activity in activating MLCK in the absence of CaM. It is strictly for research use only (RUO).
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| Molecular Formula |
C120H205F3N32O43
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| Molecular Weight |
2841.10
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| Appearance |
Solid powder
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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 Note: Please store this product in a sealed and protected environment (e.g. under nitrogen), avoid exposure to moisture and light. |
| 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 | 0.3520 mL | 1.7599 mL | 3.5198 mL | |
| 5 mM | 0.0704 mL | 0.3520 mL | 0.7040 mL | |
| 10 mM | 0.0352 mL | 0.1760 mL | 0.3520 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.
Products are for research use only; We do not sell to patients
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