Treatment of adenoid cystic carcinoma (ACC) cell lines (ACC-83 and ACC-LM) with 5 µM TNF-α (31-45), human for 1 hour activated the NF-κB pathway, as evidenced by increased phosphorylation of the P65 subunit. This activation promoted cisplatin resistance and enhanced the migration and invasion capabilities of the ACC cells. The use of TNF-α (31-45), human did not alter the mRNA or protein expression levels of RPS3. [2]

NF-κB Pathway Activation Assay: To activate the NF-κB pathway, ACC cells were treated with TNF-α (31-45), human at a final concentration of 5 µM for 1 hour. The activation status of the pathway was subsequently assessed by western blot analysis of phosphorylated P65 (P-P65) levels. [2]
Cisplatin Resistance (IC₅₀) Assay: Cells were seeded into 96-well plates at 10,000 cells/well. After treatment with TNF-α (31-45), human (5 µM, 1 hour) or other conditions, cisplatin was added at a series of concentrations (0, 2.5, 5, 10, 20, 40, 80, 160 µM) for 48 hours. Cell viability was measured using a Cell Counting Kit-8 (CCK-8) assay to determine the half-maximal inhibitory concentration (IC₅₀) of cisplatin. [2]
Migration and Invasion Assay (Transwell): For migration assays, 30,000 cells in medium containing 1% FBS were seeded into the upper chamber of a Transwell insert (8-µm pore size). For invasion assays, the upper chamber was coated with matrix gel. The lower chamber contained complete medium with 10% FBS. After pretreatment with TNF-α (31-45), human (5 µM, 1 hour), cells were allowed to migrate or invade for 24 hours. Cells on the lower surface of the membrane were fixed, stained with crystal violet, and counted under a microscope. [2]

[1]. Idriss HT, Naismith JH. TNF alpha and the TNF receptor superfamily: structure-function relationship(s). Microsc Res Tech. 2000 Aug 1;50(3):184-95.

[2]. RPS3 Promotes the Metastasis and Cisplatin Resistance of Adenoid Cystic Carcinoma. Front Oncol. 2022 Jun 30:12:804439.

[3]. The Role of Tumor Necrosis Factor Alpha (TNF-α) in Autoimmune Disease and Current TNF-α Inhibitors in Therapeutics. Int J Mol Sci. 2021 Mar 8;22(5):2719.

Tumor necrosis factor-α (TNF-α) Biology: Tumor necrosis factor-α (TNF-α) is a pleiotropic homotrimeric cytokine, primarily known as a key regulator of inflammatory responses. It exists in a soluble form (sTNF-α) and a transmembrane precursor form (tmTNF-α). Its biological activity is mediated by two receptors: TNFR1 (widely expressed, containing a death domain, primarily pro-inflammatory and pro-apoptotic) and TNFR2 (primarily expressed on immune cells, promoting cell survival, proliferation, and tissue regeneration). Dysregulation or overactivation of TNF-α signaling is associated with the pathogenesis of various autoimmune diseases. [1]
Role in autoimmune diseases: This review details the pathogenic role of excessive TNF-α in a variety of autoimmune diseases: in rheumatoid arthritis (RA), it activates synovial fibroblasts and osteoclasts, leading to joint destruction; in psoriatic arthritis (PsA) and psoriasis (PS), it promotes keratinocyte proliferation and inflammatory cell recruitment; in inflammatory bowel disease (IBD), it leads to intestinal fibrosis, matrix degradation and epithelial damage; in non-infectious uveitis (NIU), it promotes T cell differentiation and leukocyte infiltration into the eye. [1]
Therapeutic TNF-α inhibitors: This article reviews several clinically approved TNF-α biologics, but none of them are peptides “TNF-α (31-45), human”. These drugs include infliximab (a chimeric monoclonal antibody), etanercept (a TNFR2-Fc fusion protein), adalimumab (a fully human monoclonal antibody), cetozumab pegylated Fab' fragment, and golimumab (a fully human monoclonal antibody). Their mechanism of action is to block the interaction between TNF-α and its receptor (TNFR1/2). The review also lists some FDA-approved biosimilars of the original drugs and mentions novel anti-TNF drugs under development (e.g., ozolizumab). [1] Important note: This review article does not discuss, study, or mention the molecule "TNF-α (31-45), human". The article focuses on the role of endogenous TNF-α cytokines as pathogenic factors and protein-based therapeutics that block their activity. [1] In this study, human tumor necrosis factor α (31-45) was used only as a pharmacological tool to activate the NF-κB signaling pathway in vitro in order to investigate the role of this pathway in regulating cisplatin resistance and metastatic behavior in adenoid cystic cancer cells. [2]

C69H122N26O22

1667.86798

1666.92

144796-71-4

TNF-α (31-45), human TFA

71311650

White to off-white solid powder

1.697

26

26

57

117

3550

14

C[C@@H](C(=O)N[C@@H](CC(=O)N)C(=O)N[C@@H](C)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](C)C(=O)N[C@@H](CC(=O)N)C(=O)NCC(=O)N[C@@H](C(C)C)C(=O)N[C@@H](CCC(=O)O)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CCCN=C(N)N)C(=O)N[C@@H](CC(=O)O)C(=O)O)NC(=O)[C@H](CCCN=C(N)N)NC(=O)[C@H](CCCN=C(N)N)N

HYFATHUJEOQTRM-PJKQPBPGSA-N

InChI=1S/C69H122N26O22/c1-30(2)23-41(93-64(114)42(24-31(3)4)89-53(103)35(10)85-62(112)45(27-48(72)97)91-55(105)34(9)83-58(108)38(16-13-21-80-68(75)76)86-56(106)37(70)15-12-20-79-67(73)74)61(111)84-36(11)54(104)90-44(26-47(71)96)57(107)82-29-49(98)95-52(33(7)8)65(115)88-40(18-19-50(99)100)60(110)92-43(25-32(5)6)63(113)87-39(17-14-22-81-69(77)78)59(109)94-46(66(116)117)28-51(101)102/h30-46,52H,12-29,70H2,1-11H3,(H2,71,96)(H2,72,97)(H,82,107)(H,83,108)(H,84,111)(H,85,112)(H,86,106)(H,87,113)(H,88,115)(H,89,103)(H,90,104)(H,91,105)(H,92,110)(H,93,114)(H,94,109)(H,95,98)(H,99,100)(H,101,102)(H,116,117)(H4,73,74,79)(H4,75,76,80)(H4,77,78,81)/t34-,35-,36-,37-,38-,39-,40-,41-,42-,43-,44-,45-,46-,52-/m0/s1

(2S)-2-[[(2S)-2-[[(2S)-2-[[(2S)-2-[[(2S)-2-[[2-[[(2S)-4-amino-2-[[(2S)-2-[[(2S)-2-[[(2S)-2-[[(2S)-2-[[(2S)-4-amino-2-[[(2S)-2-[[(2S)-2-[[(2S)-2-amino-5-(diaminomethylideneamino)pentanoyl]amino]-5-(diaminomethylideneamino)pentanoyl]amino]propanoyl]amino]-4-oxobutanoyl]amino]propanoyl]amino]-4-methylpentanoyl]amino]-4-methylpentanoyl]amino]propanoyl]amino]-4-oxobutanoyl]amino]acetyl]amino]-3-methylbutanoyl]amino]-4-carboxybutanoyl]amino]-4-methylpentanoyl]amino]-5-(diaminomethylideneamino)pentanoyl]amino]butanedioic acid

2934.99.9001

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.

Room temperature (This product is stable at ambient temperature for a few days during ordinary shipping and time spent in Customs)

H2O : ~50 mg/mL (~29.98 mM)

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.

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Injection Formulation 1: DMSO : Tween 80： Saline = 10 : 5 : 85 (i.e. 100 μL DMSO stock solution → 50 μL Tween 80 → 850 μL Saline)
*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).

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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)]
*Preparation of 20% SBE-β-CD in Saline (4°C，1 week): Dissolve 2 g SBE-β-CD in 10 mL saline to obtain a clear solution.
Injection Formulation 5: 2-Hydroxypropyl-β-cyclodextrin : Saline = 50 : 50 (i.e. 500 μL 2-Hydroxypropyl-β-cyclodextrin → 500 μL Saline)
Injection Formulation 6: DMSO : PEG300 : castor oil : Saline = 5 : 10 : 20 : 65 (i.e. 50 μL DMSO → 100 μLPEG300 → 200 μL castor oil → 650 μL Saline)
Injection Formulation 7: Ethanol : Cremophor : Saline = 10： 10 : 80 (i.e. 100 μL Ethanol → 100 μL Cremophor → 800 μL Saline)
Injection Formulation 8: Dissolve in Cremophor/Ethanol (50 : 50), then diluted by Saline
Injection Formulation 9: EtOH : Corn oil = 10 : 90 (i.e. 100 μL EtOH → 900 μL Corn oil)
Injection Formulation 10: EtOH : PEG300：Tween 80 : Saline = 10 : 40 : 5 : 45 (i.e. 100 μL EtOH → 400 μLPEG300 → 50 μL Tween 80 → 450 μL Saline)

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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).

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Oral Formulation 3: Dissolved in PEG400
Oral Formulation 4: Suspend in 0.2% Carboxymethyl cellulose
Oral Formulation 5: Dissolve in 0.25% Tween 80 and 0.5% Carboxymethyl cellulose
Oral Formulation 6: Mixing with food powders

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Note: Please be aware that the above formulations are for reference only. InvivoChem strongly recommends customers to read literature methods/protocols carefully before determining which formulation you should use for in vivo studies, as different compounds have different solubility properties and have to be formulated differently.

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 (Please use freshly prepared in vivo formulations for optimal results.)