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| 1mg |
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| Targets |
TRAF6 peptide TFA specifically targets the interaction between two key proteins involved in cellular signaling: TRAF6 (TNF receptor-associated factor 6) and p62 (sequestosome-1). By binding to this interface, it acts as a selective inhibitor of the TRAF6-p62 complex formation, preventing TRAF6-mediated ubiquitination of its substrates. A key mechanism of this inhibitor is its ability to eliminate NGF-dependent TrkA ubiquitination, which is a critical step in the signaling pathway for neurotrophins like NGF. Thus, the target pathway is the TRAF6-p62 axis, which has been implicated in neuroinflammation and protein aggregation in several neurological disorders.
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| ln Vitro |
In vitro, TRAF6 peptide TFA has demonstrated its ability to block specific signaling pathways. It has been shown to abolish NGF (Nerve Growth Factor)-dependent ubiquitination of the TrkA receptor [28L12-L13]. This inhibition of TrkA ubiquitination disrupts downstream signaling cascades. The peptide is cell-permeable, making it effective in cell culture models. A concentration of 100 mg/mL is soluble in water (with ultrasonic assistance), allowing for easy handling in aqueous buffers for in vitro experiments. The specific IC50 is not reported for the peptide alone, as its function is to disrupt a protein-protein interaction.
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| ln Vivo |
While in vivo activity is primarily studied in animal models of neurological diseases, the mechanism has been validated by showing that the peptide can protect cells from various pathological insults. In a cellular context, by disrupting TRAF6-p62 binding, the peptide is expected to reduce neuroinflammation, protein aggregation, and neuronal cell death. It has good research potential in a variety of neurological disease models, including Alzheimer's disease (AD), Parkinson's disease (PD), amyotrophic lateral sclerosis (ALS), traumatic brain injury (TBI), epilepsy, and stroke, where the TRAF6-p62 interaction is believed to play a pathogenic role.
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| Enzyme Assay |
The TRAF6-p62 interaction can be studied in vitro using a solid-phase binding assay or a pull-down assay. To perform a binding assay, recombinant TRAF6 protein is immobilized on a 96-well plate. Increasing concentrations of the TRAF6 peptide TFA are incubated with a labeled p62 protein (e.g., biotinylated p62). After washing steps to remove unbound p62, the bound p62 is detected using a streptavidin-conjugated HRP and a colorimetric substrate. The ability of the peptide to inhibit this interaction, as measured by a decrease in signal, demonstrates its potency.
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| Cell Assay |
For in vitro cellular assays, cells (e.g., neuronal cell lines, HeLa cells) are cultured in appropriate medium. To assess the mechanism, cells can be treated with NGF to stimulate TrkA signaling, and then incubated with various concentrations of TRAF6 peptide TFA (e.g., 1-50 microM) for 4-24 hours. The cells are then lysed, and the level of TrkA ubiquitination is analyzed by immunoprecipitation with a TrkA antibody followed by western blotting with a ubiquitin antibody. Alternatively, the effect on cell viability in a disease model can be measured by treating cells with an insult like amyloid-beta or MPP+ (a Parkinson's model) with or without the peptide, followed by an MTT or LDH assay.
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| Animal Protocol |
In vivo animal studies are typically performed in rodent models of the disease of interest. For a model of Parkinson's disease, mice might be injected with MPTP (a neurotoxin) to induce dopaminergic neuron loss and motor deficits. The TRAF6 peptide TFA can be administered via intraperitoneal (IP) injection at a dose of 1-10 mg/kg in saline, either daily or every other day, for a period of 1-4 weeks. Behavioral tests (e.g., rotarod, open field test) are performed to assess motor function. At the end of the study, the mice are sacrificed, and brain tissue (e.g., substantia nigra) is collected to measure levels of inflammatory cytokines (IL-6, TNF-alpha) and aggregation-prone proteins (e.g., alpha-synuclein) by ELISA or western blot. The protective effect of the peptide is evaluated by comparing these parameters to a control group.
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| ADME/Pharmacokinetics |
As a peptide, the pharmacokinetic properties of TRAF6 peptide TFA are not fully characterized and are not the primary focus of its research use, as it is a tool compound. However, it is designed to be cell-permeable, which is critical for its in vivo efficacy. This suggests that the peptide is able to cross cellular membranes, a property that is essential for reaching its intracellular targets (TRAF6 and p62). Pharmacokinetic studies would involve the administration of the peptide and subsequent measurement of its concentration in plasma and tissues (e.g., brain) using LC-MS/MS to assess its absorption, distribution, and clearance, which is generally rapid for peptides.
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| Toxicity/Toxicokinetics |
Specific toxicity data for TRAF6 peptide TFA is not publicly available. As a research peptide, its toxicological profile has not been extensively studied beyond its use in animal models. However, it has been used in animal models of disease, suggesting that at the concentrations and dosing regimens used for research (e.g., 1-10 mg/kg IP), it is generally well-tolerated, with no reports of acute toxicity. As with all research chemicals, standard safety practices (gloves, lab coat, eye protection) should be followed. It is strictly for research purposes only and not for human or clinical use.
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| References | |
| Additional Infomation |
TRAF6 peptide TFA is a cell-permeable peptide inhibitor that specifically disrupts the TRAF6-p62 interaction, effectively blocking NGF-induced TrkA ubiquitination. This makes it a valuable tool for studying the mechanisms of neurotrophic signaling and protein aggregation in a variety of neurological conditions, including Alzheimer's, Parkinson's, and ALS. The peptide is supplied as a TFA salt to improve its solubility and stability. It is an important research tool for understanding the role of the TRAF6-p62 pathway in both neurobiology and inflammation. It is strictly for research use and has not been approved for clinical use.
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| Molecular Formula |
C145H238N34O44.XC2HF3O2
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| Molecular Weight |
3161.64 (free acid)
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| Related CAS # |
TRAF6 peptide;852805-92-6
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| Appearance |
White to off-white 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) |
H2O :~100 mg/mL
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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.) |
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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