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| Targets |
TNF-alpha (46-65), human TFA is a peptide fragment of the full-length TNF-alpha protein. Its intended target is not a single enzyme or receptor but rather the binding interfaces on the TNF-alpha protein itself. This fragment is derived from a specific region of the cytokine and can be used to study how TNF-alpha interacts with its receptors (TNFR1 and TNFR2), which are part of the TNF receptor family and the Apoptosis pathway [32L6-L7]. By blocking or mimicking the activity of the full-length cytokine, this peptide is a useful tool for mapping functional epitopes and exploring cytokine-receptor interactions.
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| ln Vitro |
The peptide fragment itself is not known to possess the full functional activity (e.g., inducing apoptosis or inflammation) of the mature TNF-alpha cytokine. Instead, it is used as an analytical tool and an immunogen to study the properties of the full-length protein. In vitro activity is therefore characterized by its ability to bind to antibodies raised against this region of TNF-alpha, which can be measured using an enzyme-linked immunosorbent assay (ELISA). It can also be used in competitive binding assays to study the binding of larger, active TNF-alpha fragments or the whole cytokine to its receptor.
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| ln Vivo |
In vivo activity data is not generally applicable to a short peptide fragment like TNF-alpha (46-65). Its primary use in a living system would be as an immunogen or an antigen to generate antibodies for research purposes. As a fragment, it is not expected to induce the robust inflammatory or apoptotic signaling characteristic of full-length TNF-alpha. The physiological effect of TNF-alpha, a proinflammatory cytokine, includes inducing necrosis or apoptosis, activating the NF-kB pathway via TNFR2, and promoting cancer growth and metastasis. However, this short peptide fragment is not used to study these effects directly.
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| Enzyme Assay |
The binding of TNF-alpha (46-65), human TFA to its target protein is typically studied using a solid-phase binding assay, such as an enzyme-linked immunosorbent assay (ELISA). In one protocol, a known TNF-alpha antibody (that recognizes this epitope) is coated onto a 96-well ELISA plate. After blocking, the peptide is added at increasing concentrations. Bound peptide is then detected using a biotinylated secondary antibody (if the primary is not labeled) or a specific detection antibody, followed by streptavidin-HRP and a colorimetric substrate. The binding affinity of the peptide for the antibody can be determined by measuring the half-maximal effective concentration (EC50) for binding.
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| Cell Assay |
For in vitro cellular assays, a peptide fragment such as TNF-alpha (46-65), human TFA can be used to study its effect on cultured immune cells. A typical protocol involves culturing human peripheral blood mononuclear cells (PBMCs) or specific cell lines (e.g., THP-1 monocytes). The cells are treated with various concentrations of the peptide (e.g., 0.1-100 microg/mL) for 6-24 hours. After incubation, the culture supernatant is collected, and the secretion of other cytokines (e.g., IL-6, IL-1beta) is measured by ELISA to assess any immunomodulatory effects. Alternatively, the peptide can be used to block the activity of full-length TNF-alpha by pre-incubating it with the cells before adding the cytokine.
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| Animal Protocol |
Since TNF-alpha (46-65), human TFA is not a therapeutic drug but a research reagent, it is not typically used in in vivo animal studies as an active therapeutic agent. Its most common in vivo use would be as an immunogen to generate antibodies. A standard protocol for this involves emulsifying 50-200 microg of the peptide in complete Freund's adjuvant (CFA) and injecting it subcutaneously into rabbits or mice. This is followed by 2-3 booster injections in incomplete Freund's adjuvant (IFA) at 2-3 week intervals. Serum is collected from the animals to isolate polyclonal antibodies against the TNF-alpha (46-65) epitope, which can then be used in various assays.
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| ADME/Pharmacokinetics |
The pharmacokinetic properties of TNF-alpha (46-65), human TFA are not relevant to its use as a research peptide. It is not intended for use as a therapeutic agent, and its absorption, distribution, metabolism, and excretion (ADME) are not typically characterized. As a short peptide, it is likely to be rapidly degraded in vivo by proteases if it were to be administered. Its utility lies in its use as a soluble research tool. The TFA salt form is used to enhance the solubility and stability of the peptide in aqueous buffers for laboratory handling, facilitating its use in immunological and biochemical assays.
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| Toxicity/Toxicokinetics |
Specific toxicity data for TNF-alpha (46-65), human TFA is not available. As a short peptide fragment of a human cytokine, it is generally considered to have low toxicity at the concentrations used in laboratory research (e.g., microg to mg/mL). However, as with all research chemicals, standard laboratory safety practices should be followed, including the use of gloves and a lab coat to avoid skin contact or inhalation. The peptide is strictly for research use only and is not intended for human or clinical use. It should be stored as a powder at -80degC for long-term stability [32L8].
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| Additional Infomation |
TNF-alpha (46-65), human TFA (TFA salt) is a peptide fragment of human TNF-alpha. It is a valuable research tool for the biochemical, immunological, and therapeutic research of cytokine functions. The TFA salt form improves its solubility and stability. The peptide is derived from a region of the TNF-alpha protein that is important for receptor binding, making it a useful tool for studying cytokine-receptor interactions. This peptide is strictly for research use only. It is not a therapeutic drug, has not been approved by the FDA, and is not intended for clinical use. It is typically stored as a dry powder at -80degC to ensure its long-term stability and prevent degradation [32L8].
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| Molecular Formula |
C112H173F3N24O32
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| Molecular Weight |
2424.71
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| Related CAS # |
TNF-α (46-65), human;144796-72-5
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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 | 0.4124 mL | 2.0621 mL | 4.1242 mL | |
| 5 mM | 0.0825 mL | 0.4124 mL | 0.8248 mL | |
| 10 mM | 0.0412 mL | 0.2062 mL | 0.4124 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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