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| Targets |
This peptide fragment specifically targets the TNF receptor (TNFR) signaling pathway, which ultimately leads to the activation of the NF-kappaB transcription factor. The parent molecule, TNF-alpha, is a pro-inflammatory cytokine that binds to its receptors (TNFR1 and TNFR2). This binding recruits adapter proteins and triggers a signaling cascade that leads to the activation of NF-kappaB. TNF-alpha (31-45), human TFA acts as an activator of this pathway, and the TNF receptor and NF-kB are its primary targets within the Apoptosis and NF-kB pathways [33L5-L6].
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| ln Vitro |
In vitro, TNF-alpha (31-45), human TFA has been shown to be a potent activator of the NF-kappaB pathway. In a cellular context, it has been demonstrated to promote cisplatin resistance, an effect that is mediated by the activation of the NF-kappaB pathway. At a concentration of 5 microM, treatment for 1 hour was sufficient to promote this effect, while it did not alter RPS3 expression levels, suggesting a specific mechanism of action [33L13-L15]. This activity makes it a valuable tool for studying the link between inflammation, NF-kappaB activation, and drug resistance in cancer cells.
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| ln Vivo |
In vivo activity data is not generally reported for this specific peptide fragment. As a fragment of the TNF-alpha cytokine, it is not typically used in animal studies as a direct therapeutic agent. However, the full-length TNF-alpha has profound in vivo effects, including promoting tumor growth, invasion, and metastasis by stimulating the NF-kappaB pathway through its interaction with TNFR2 [33L11-L12]. This peptide fragment is a research tool for studying the mechanism of these processes at a molecular level in vitro.
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| Enzyme Assay |
The activity of TNF-alpha (31-45), human TFA is characterized by its ability to bind to its target receptors. A standard cell-free assay to study this is an enzyme-linked immunosorbent assay (ELISA). A 96-well plate can be coated with recombinant TNF receptor (TNFR2) protein. The plate is blocked with a protein solution (e.g., BSA) to prevent non-specific binding. Increasing concentrations of the TNF-alpha (31-45) peptide are added to the wells and allowed to bind. Bound peptide is then detected using a specific primary antibody against this TNF-alpha epitope, followed by an HRP-conjugated secondary antibody. The signal is developed using a colorimetric substrate and read on a plate reader to determine the binding affinity.
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| Cell Assay |
For in vitro cellular assays, cultured cells that are responsive to TNF-alpha (e.g., HeLa, HEK-293, or cancer cell lines) can be used. Cells are plated in a 96-well plate and allowed to attach. They are then treated with the TNF-alpha (31-45) peptide at varying concentrations (e.g., 0.1-100 microM) for 1-24 hours. After treatment, the cells are lysed, and the level of NF-kappaB activation is measured. This can be done using a luciferase reporter assay (if cells are transfected with an NF-kappaB-responsive luciferase construct), or more commonly by preparing nuclear extracts and performing an NF-kappaB ELISA-based transcription factor assay kit to detect p65 binding to its DNA consensus sequence.
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| Animal Protocol |
The TNF-alpha (31-45), human TFA peptide is not intended for use as a therapeutic agent in animal studies, and its in vivo use would be primarily as an antigen. A common animal protocol is the generation of anti-TNF-alpha antibodies. Peptide (100-200 microg) is emulsified in Complete Freund's Adjuvant (CFA) and injected subcutaneously into a rabbit or mouse. Booster immunizations are given 2-4 weeks later using the same amount of peptide emulsified in Incomplete Freund's Adjuvant (IFA). After 2-3 boosters, blood is collected, and serum is tested for antibody titer by ELISA. This polyclonal antibody can then be used in various immunological techniques to detect human TNF-alpha.
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| ADME/Pharmacokinetics |
The pharmacokinetic properties of TNF-alpha (31-45), human TFA are not characterized, as it is a research peptide and not a therapeutic drug candidate. As a small peptide, if it were to enter the bloodstream, it would be rapidly degraded and cleared. The TFA salt is a common counterion used during peptide purification (HPLC). Its presence is known to affect the peptide's net weight, appearance, and solubility. The TFA salt contributes to the total mass of the product, which must be accounted for when preparing solutions. The compound is generally soluble in water, and the TFA salt helps to enhance its solubility and stability during storage.
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| Toxicity/Toxicokinetics |
There is no specific toxicity data available for TNF-alpha (31-45), human TFA. As a short peptide fragment of a human cytokine, it is generally considered to have low toxicity for laboratory use. However, as with all research chemicals, standard safety practices should be followed, including wearing gloves and a lab coat to prevent skin contact or accidental inhalation. The peptide should be handled in a well-ventilated area. It is strictly for research use only and is not intended for diagnostic, therapeutic, or clinical applications. The free base form is typically stored at -20degC for long-term stability [33L7-L8].
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| References |
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| Additional Infomation |
TNF-alpha (31-45), human TFA is a peptide fragment of the human tumor necrosis factor-alpha (TNF-alpha) protein. Its sequence is RRANALLANGVELRD [33L5]. It is supplied as a TFA salt, which is a common salt form for synthetic peptides used in research. This peptide is a valuable tool for studying the role of TNF-alpha in activating the NF-kB pathway, a critical signaling pathway involved in inflammation, immunity, cell survival, and oncogenesis. It is strictly for research use only and is not a therapeutic drug, not for clinical use, and not approved by any regulatory agency. It is typically stored as a powder at -80degC to ensure long-term stability [33L7].
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| Molecular Formula |
C71H123F3N26O24
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| Molecular Weight |
1781.89
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| Related CAS # |
TNF-α (31-45), human;144796-71-4
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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 | 0.5612 mL | 2.8060 mL | 5.6120 mL | |
| 5 mM | 0.1122 mL | 0.5612 mL | 1.1224 mL | |
| 10 mM | 0.0561 mL | 0.2806 mL | 0.5612 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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