TPCK
Based on 1 publication(s) in Google Scholar
TPCK (L-1-Tosylamido-2-phenylethyl chloromethyl ketone; L-TPCK) is an effective serine protease inhibitor and also a blocker of the PDK1/Akt pathway. TPCK can modify the E7 protein in actively keratinocyte cells. TPCK can induce cellular apoptosis, suppress tumor growth, reduce hypoxic-ischemic brain injury in rat pups, and affect vascular permeability in inflamed rats.
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研究用途以外に使用した場合、当社は一切の責任を負いかねます。
- 純度 : 98.05%
- CAS 番号: 402-71-1
- 分子式: C17H18ClNO3S
- 分子量:351.85
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保管条件:Powder -20°C, 3 years , 4°C, 2 years ; In solvent -80°C, 6 months , -20°C, 1 month
MedChemExpress(MCE)の使用を引用している文献 TPCK
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生物活性
製品説明
Cellular Effect
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Cell Line
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Type | Value | Description | References |
|---|---|---|---|---|
| HEK293 | IC50 |
10.8 μM
Compound: TPCK
|
Inhibition of TNFalpha-induced NFkappaB (unknown origin) activation expressed in HEK293 cells after 6 hrs by luciferase reporter gene assay
Inhibition of TNFalpha-induced NFkappaB (unknown origin) activation expressed in HEK293 cells after 6 hrs by luciferase reporter gene assay
|
[PMID: 23316950] |
| HEK293 | IC50 |
11.1 μM
Compound: TPCK
|
Cytotoxicity against HEK293 cells
Cytotoxicity against HEK293 cells
|
[PMID: 24533857] |
| HEK293 | IC50 |
3.76 μM
Compound: TPCK
|
Inhibition of TNF-alpha-induced NFkappaB activation in human HEK293 cells after 6 hrs by luciferase reporter gene assay
Inhibition of TNF-alpha-induced NFkappaB activation in human HEK293 cells after 6 hrs by luciferase reporter gene assay
|
[PMID: 21261296] |
| HEK293 | IC50 |
3.8 μM
Compound: TPCK
|
Inhibition of TNFalpha-induced NF-kappaB activity expressed in human HEK 293 cells after 6 hrs by luciferase reporter gene assay
Inhibition of TNFalpha-induced NF-kappaB activity expressed in human HEK 293 cells after 6 hrs by luciferase reporter gene assay
|
[PMID: 22850207] |
| HEK293 | IC50 |
3.8 μM
Compound: TPCK
|
Cytotoxicity against human HEK293 cells assessed as cell survival by SRB assay
Cytotoxicity against human HEK293 cells assessed as cell survival by SRB assay
|
[PMID: 23316950] |
| HEK293 | IC50 |
3.8 μM
Compound: TPCK
|
Inhibition of TNFalpha-induced nuclear factor-kappa-B activation in human HEK293 cells at by luciferase reporter gene assay
Inhibition of TNFalpha-induced nuclear factor-kappa-B activation in human HEK293 cells at by luciferase reporter gene assay
|
[PMID: 23994869] |
| HEK293 | IC50 |
3.8 μM
Compound: TPCK
|
Inhibition of TNF-alpha-induced NF-kappaB activity in HEK293 cells after 6 hrs by luciferase reporter gene assay
Inhibition of TNF-alpha-induced NF-kappaB activity in HEK293 cells after 6 hrs by luciferase reporter gene assay
|
[PMID: 24533857] |
| HEK293 | IC50 |
3.8 μM
Compound: TPCK
|
Inhibition of TNF-alpha-induced NF-kappaB activity in HEK293 cells after 6 hrs by luciferase reporter gene assay
Inhibition of TNF-alpha-induced NF-kappaB activity in HEK293 cells after 6 hrs by luciferase reporter gene assay
|
[PMID: 24992702] |
| HEK293 | IC50 |
3.8 μM
Compound: TPCK
|
Inhibition of TNFalpha-induced NF-kappaB activity (unknown origin) transfected in HEK293 cells after 6 hrs by luciferase reporter gene assay
Inhibition of TNFalpha-induced NF-kappaB activity (unknown origin) transfected in HEK293 cells after 6 hrs by luciferase reporter gene assay
|
[PMID: 26343828] |
| HEK293 | IC50 |
3.8 μM
Compound: TPCK
|
Inhibition of TNF-alpha-activated NFkappaB (unknown origin) transfected in HEK293 cells after 6 hrs by luciferase reporter gene assay
Inhibition of TNF-alpha-activated NFkappaB (unknown origin) transfected in HEK293 cells after 6 hrs by luciferase reporter gene assay
|
[PMID: 27196335] |
| HEK293 | IC50 |
4.9 μM
Compound: N-tosyl-L-phenylalanine-chloromethyl ketone
|
Inhibition of TNF-alpha-induced NFkappaB activation in human HEK293 cells after 6 hrs by luciferase assay
Inhibition of TNF-alpha-induced NFkappaB activation in human HEK293 cells after 6 hrs by luciferase assay
|
[PMID: 21978950] |
| HEK293 | IC50 |
5.05 μM
Compound: TPCK
|
Inhibition of TNF-alpha activated nuclear factor-kappa B in HEK293 cells after 6 hrs by luciferase reporter gene assay
Inhibition of TNF-alpha activated nuclear factor-kappa B in HEK293 cells after 6 hrs by luciferase reporter gene assay
|
[PMID: 26048799] |
| HEK293 | IC50 |
5.09 μM
Compound: TPCK
|
Inhibition of NF-kappaB expressed in human HEK293 cells by luciferase reporter gene based luminometric analysis
Inhibition of NF-kappaB expressed in human HEK293 cells by luciferase reporter gene based luminometric analysis
|
[PMID: 21105712] |
| HEK293 | IC50 |
5.09 μM
Compound: TPCK
|
Inhibition of TNFalpha-activated NF-kappaB expressed in HEK293 cells by luciferase reporter gene assay
Inhibition of TNFalpha-activated NF-kappaB expressed in HEK293 cells by luciferase reporter gene assay
|
[PMID: 22115839] |
| HEK293 | IC50 |
5.09 μM
Compound: TPCK
|
Inhibition of TNF-alpha activated NF-kappaB expressed in HEK293 cells by luciferase reporter gene assay
Inhibition of TNF-alpha activated NF-kappaB expressed in HEK293 cells by luciferase reporter gene assay
|
[PMID: 22386564] |
| HEK293 | IC50 |
5.09 μM
Compound: TPCK
|
Inhibition of TNFalpha-induced NF-kappaB activity expressed in HEK293 cells by luciferase reporter gene assay
Inhibition of TNFalpha-induced NF-kappaB activity expressed in HEK293 cells by luciferase reporter gene assay
|
[PMID: 23142320] |
| HEK293 | IC50 |
5.09 μM
Compound: TPCK
|
Inhibition of TNF-alpha induced NF-kappaB transcriptional activity in HEK293 cells expressing NF-kappaB-Luc measured after 48 hrs by luciferase assay
Inhibition of TNF-alpha induced NF-kappaB transcriptional activity in HEK293 cells expressing NF-kappaB-Luc measured after 48 hrs by luciferase assay
|
[PMID: 27155469] |
| HEK293 | IC50 |
5.09 μM
Compound: TPCK
|
Inhibition of TNFalpha-activated nuclear factor-kappa B activity (unknown origin) expressed in HEK293 cells incubated for 6 hrs by luciferase reporter gene based assay
Inhibition of TNFalpha-activated nuclear factor-kappa B activity (unknown origin) expressed in HEK293 cells incubated for 6 hrs by luciferase reporter gene based assay
|
[PMID: 27210437] |
| HEK293 | IC50 |
5.1 μM
Compound: TPCK
|
Inhibition of TNFalpha-induced human NFkappaB activity in HEK293 cells incubated for 6 hrs followed by compound wash out measured after 5 mins by by luciferase assay
Inhibition of TNFalpha-induced human NFkappaB activity in HEK293 cells incubated for 6 hrs followed by compound wash out measured after 5 mins by by luciferase assay
|
[PMID: 22712432] |
| HEK293 | IC50 |
5.3 μM
Compound: TPCK
|
Inhibition of TNFalpha-induced NFkappaB activation in human HEK293 cells after 6 hrs by luciferase assay
Inhibition of TNFalpha-induced NFkappaB activation in human HEK293 cells after 6 hrs by luciferase assay
|
[PMID: 32163285] |
体外実験
TPCK shows slight toxicity to mammalian cells (CC50 = 138.8 µM), but it significantly inhibits the pre-flagellate morphology of L. amazonensis PH8 and Josefa strains (IC50 values of 14.6 µM and 31.7 µM, respectively), with an IC50 value of 11.3 µM for the pre-flagellate morphology of L. infantum. Additionally, TPCK is also effective against the intracellular amastigote form, showing IC50 values of 14.2 µM and 16.6 µM for L. amazonensis PH8 and Josefa strains, and 21.7 µM for the amastigote form of L. infantum[2].
TPCK (40 μM, 1 h) enhances wortmannin-dependent caspase activity in LNCaP cells but inhibits TRAIL-dependent caspase activity[3].
TPCK (40 μM, 1 h) inhibits the formation of TRAIL-DISC in PC3 cells but does not inhibit the formation of Fas-DISC and reduces caspase2 levels[3].
TPCK (0-40 μM, 24 h) kills PC3 cells in a dose-dependent manner, inducing apoptosis[3].
TPCK (40 μM, 3 h) reduces the level of BAD-ser136 in LNCaP cells and eliminates AR in the cytoplasm and granules[3].
MedChemExpress (MCE) has not independently confirmed the accuracy of these methods. They are for reference only. Further protocols information, click here.
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Cell Line:PC3; LNCaP
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Concentration:40 μM
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Incubation Time:1 h; 3 h
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Result:Reduced the level of caspase-2 and inhibited TRAIL-induced activation of caspase-3, -7, -8, and -9. Did not reduce BAD levels, but reduced the level of BAD-ser136 and eliminated AR in the cytosol and granules.
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Cell Line:PC3
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Concentration:0, 2.5, 5, 10, 20, 40 μM
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Incubation Time:24 h
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Result:Killed PC3 cells in a dose-dependent manner.
体内実験
TPCK (5-100 mg/kg, intraperitoneal injection, single dose) diminishes DNA fragmentation, nitric oxide production, and brain damage in a neonatal mouse model of hypoxic-ischemic brain injury[4].
TPCK (0.5-20 mg/kg, injected into the air sac, single dose) increases vascular permeability in inflamed rats, which then decline[5].
MedChemExpress (MCE) has not independently confirmed the accuracy of these methods. They are for reference only.
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Animal Model:BALB/c mice infected with L. amazonensis [2]
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Dosage:15, 30, 45, 60 mg/kg; three times a week; 10 times
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Administration:Intraperitoneal injection (i.p.)
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Result:Reduced lesion size and parasite burden in foot pads and spleen.
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Animal Model:7-day-old rats with right carotid artery ligation[4]
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Dosage:5, 10, 20, 50, 100 mg/kg; single dose
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Administration:Intraperitoneal injection (i.p.)
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Result:Reduced the weight of the right hemisphere of the brain, reduced the number of DNA breaks in cells, abolished the increase in nitrate/nitrite metabolites within 6 hours after injury, and reduced DNA fragmentation, nitric oxide production, and brain damage.
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Animal Model:Carrageenan-air sac inflammation rats[5]
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Dosage:0.5, 4, 20 mg/kg; single dose
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Administration:Injection into the air sac
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Result:Showed that vascular permeability increased rapidly and then gradually decreased.
化学情報
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CAS 番号 402-71-1
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性状 Solid
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分子量 351.85
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分子式 C17H18ClNO3S
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Color White to off-white
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SMILES
CC1=CC=C(C=C1)S(N[C@@H](CC2=CC=CC=C2)C(CCl)=O)(=O)=O
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別名
L-1-Tosylamido-2-phenylethyl chloromethyl ketone; L-TPCK
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輸送条件
Room temperature in continental US; may vary elsewhere.
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保管条件
Powder -20°C 3 years 4°C 2 years In solvent -80°C 6 months -20°C 1 month
Publications (1)
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Journal Impact Factor
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Most Recent
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Int J Biol Macromol
Evaluation and mechanism of immune enhancement effects of Pleurotus ferulae polysaccharides-gold nanoparticles. [Abstract]2023 Feb 1:227:1015-1026. PMID: 36460244
溶剤 & 溶解度
体外:
DMSO : 100 mg/mL (284.21 mM; Need ultrasonic; Hygroscopic DMSO has a significant impact on the solubility of product, please use newly opened DMSO)
Please refer to the solubility information to select the appropriate solvent. Once prepared, please aliquot and store the solution to prevent product inactivation from repeated freeze-thaw cycles.
Storage method and period of stock solution: -80°C, 6 months; -20°C, 1 month. When stored at -80°C, please use it within 6 months. When stored at -20°C, please use it within 1 month.
Please refer to the solubility information to select the appropriate solvent. Once prepared, please aliquot and store the solution to prevent product inactivation from repeated freeze-thaw cycles.
Storage method and period of stock solution: -80°C, 6 months; -20°C, 1 month. When stored at -80°C, please use it within 6 months. When stored at -20°C, please use it within 1 month.
濃度 (開始) × 体積 (開始) = 濃度 (終了) × 体積 (終了)
体内:
Select the appropriate dissolution method based on your experimental animal and administration route.
- For the following dissolution methods, please ensure to first prepare a clear stock solution using an In Vitro approach and then sequentially add co-solvents:
- To ensure reliable experimental results, the clarified stock solution can be appropriately stored based on storage conditions. As for the working solution for In Vivo experiments, it is recommended to prepare freshly and use it on the same day.
- The percentages shown for the solvents indicate their volumetric ratio in the final prepared solution. If precipitation or phase separation occurs during preparation, heat and/or sonication can be used to aid dissolution.
Add each solvent one by one: 10% DMSO 40% PEG300 5% Tween-80 45% Saline
Solubility: ≥ 2.5 mg/mL (7.11 mM); Clear solution
This protocol yields a clear solution of ≥ 2.5 mg/mL (saturation unknown).
Taking 1 mL working solution as an example, add 100 μL DMSO stock solution (25.0 mg/mL) to 400 μL PEG300, and mix evenly; then add 50 μL Tween-80 and mix evenly; then add 450 μL Saline to adjust the volume to 1 mL.
Preparation of Saline: Dissolve 0.9 g sodium chloride in ddH₂O and dilute to 100 mL to obtain a clear Saline solution.
In Vivo Dissolution Calculator
Please enter the basic information of animal experiments:
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Recommended: Prepare an additional quantity of animals to account for potential losses during experiments.
Please enter your animal formula composition:
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%DMSO +
Recommended: Keep the proportion of DMSO in working solution below 2% if your animal is weak.
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%+
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+%Tween-80 + +
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%Saline +
The co-solvents required include: DMSO, . All of co-solvents are available by MedChemExpress (MCE). , Tween 80. All of co-solvents are available by MedChemExpress (MCE).
Working solution concentration: 0.22 mg/mL
Method for preparing stock solution: mg drug dissolved in μL DMSO. Stock solution concentration: mg/mL.
1. Take μL DMSO stock solution;
2. Add μL .
μL , mix evenly;
3. Then add μL Tween 80, mix evenly;
4. Then add μL
Please ensure that the stock solution in the first step is dissolved to a clear state, and add co-solvents in sequence. You can use ultrasonic heating (ultrasonic cleaner, recommended frequency 20-40 kHz), vortexing, etc. to assist dissolution.
プロトコル
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Apoptosis
Apoptosis, also called programmed cell death, is generally characterized by distinct morphological characteristics.
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TUNEL staining for apoptotic DNA fragmentation
TUNEL staining detects DNA strand breaks by using terminal deoxynucleotidyl transferase to add labeled nucleotides to exposed 3′-OH DNA termini, generating either microscopic staining in fixed cells or tissue sections, or fluorescence/cytometric signal in cell suspensions. TUNEL positivity reflects DNA fragmentation but should not be interpreted alone as definitive apoptosis, because TUNEL can also label necrotic, autolytic, mechanically damaged, or DNA-repair-associated DNA breaks.
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Annexin V plus membrane-impermeant dye apoptosis staining
Annexin V-based apoptosis assays rely on the detection of phosphatidylserine (PS) externalization from the inner leaflet of the plasma membrane to the outer leaflet, an early biochemical hallmark of apoptosis. Fluorescently labeled Annexin V binds PS in a calcium-dependent manner, enabling identification of early apoptotic cells by flow cytometry or fluorescence microscopy. When combined with a membrane-impermeant DNA-binding dye (e. g. , propidium iodide), this approach allows discrimination between viable (Annexin V−/dye−), early apoptotic (Annexin V+/dye−), and late apoptotic or necrotic (Annexin V+/dye+) cell populations by assessing membrane integrity and PS exposure.
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Patient-Derived Xenograft (PDX)
Patient-derived xenograft (PDX) models are generated by engrafting primary human tumor tissue directly into immunodeficient mice, allowing in vivo propagation of patient tumor biology without initial in vitro adaptation. These models are used to preserve key histopathological and molecular characteristics of the original tumor and enable assessment of tumor growth dynamics and therapeutic response in a living organism. The biological readout is tumor engraftment and subsequent growth in the murine host, which reflects the ability of human tumor cells to survive, vascularize, and expand in an immunocompromised microenvironment.
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Apoptosis Solutions
Apoptosis is a regulated, generally non-lytic cell-death pathway that removes unwanted, damaged, infected, or abnormal cells through coordinated morphological changes, caspase activation, DNA fragmentation, and membrane remodeling. The intrinsic apoptosis pathway is controlled mainly by mitochondrial outer membrane permeabilization, BCL-2 family proteins, cytochrome c release, apoptosome formation, caspase-9 activation, and downstream executioner caspase-3/7 activation. The extrinsic apoptosis pathway is initiated by death receptors such as Fas, TNFR, and TRAIL receptors, which recruit adaptor proteins and activate caspase-8 before engaging executioner caspases or mitochondrial amplification through BID cleavage. Apoptosis is linked to many phenotypes, including cancer cell killing, tissue homeostasis, immune regulation, neurodegeneration, infection response, and treatment-induced cytotoxicity; unresolved questions include how apoptosis interacts with necroptosis, pyroptosis, ferroptos
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Patient-Derived Orthotopic Xenograft (PDOX)
Patient-derived orthotopic xenograft (PDOX) modeling implants fresh patient tumor tissue or patient-derived tumor cells into the anatomically corresponding organ or tissue site of immunodeficient mice, usually by surgical orthotopic implantation, to preserve patient tumor histology, local microenvironmental context, invasion, metastatic behavior, and treatment-response features better than subcutaneous implantation. PDOX readouts include tumor engraftment, orthotopic tumor growth, local invasion, metastasis, recurrence after resection, histologic similarity to the donor tumor, biomarker retention, molecular concordance, survival, and response or resistance to therapy. PDOX models are used for preclinical drug testing and individualized therapy evaluation, but engraftment success varies by tumor type and specimen quality.
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Transepithelial/transendothelial electrical resistance assay
TEER measures electrical resistance across epithelial or endothelial monolayers cultured on permeable supports, and the readout reflects ionic conductance through the cell barrier, especially the paracellular pathway regulated by junctional integrity. TEER can be measured without destroying the monolayer and is commonly used before or during transport, permeability, barrier-disruption, and barrier-maturation experiments. TEER values are influenced by biological maturation and technical conditions; reported factors include temperature, medium formulation, passage number, electrode geometry, membrane properties, and junctional length during early monolayer maturation. Therefore, TEER should be interpreted with blank-insert subtraction, area normalization, repeated readings, and, when possible, orthogonal barrier readouts such as FITC-dextran flux or tight-junction staining.
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Subcutaneous Cell-Line-Derived Xenograft
Subcutaneous cell-line-derived xenograft (CDX) models are established by implanting cultured human cancer cell lines into immunodeficient mice, where the injected cells form localized tumors that can be monitored in vivo as a measure of tumorigenic potential, growth kinetics, and treatment response. These models are widely used in oncology research because they allow reproducible tumor formation and enable comparative assessment of tumor growth between different cell lines or genetic manipulations in a controlled in vivo microenvironment. Subcutaneous implantation of cancer cells in immunodeficient mice is a standard approach for evaluating tumor growth behavior and therapeutic response across multiple cancer types, including prostate, esophageal, pancreatic, and colon cancer models.
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Orthotopic Cell-Line Xenograft
Orthotopic cell-line xenograft models involve implantation of human cancer cell lines into the anatomically corresponding organ of immunodeficient mice to reproduce tumor growth within a native microenvironment, enabling more clinically relevant tumor behavior compared with subcutaneous models. These models are widely used because orthotopic placement better recapitulates tumor progression, including invasion and metastatic spread, which are often underrepresented in heterotopic implantation systems. Compared with conventional xenografts, orthotopic implantation is described as more technically complex but provides improved simulation of tumor-microenvironment interactions and metastatic behavior, making it particularly valuable for translational oncology research. Surgical orthotopic implantation approaches have been emphasized as enabling faithful reproduction of clinical cancer features, including metastasis and disease progression patterns that align with the tumor’s organ of origi
純度とドキュメンテーション
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取扱説明書 (2659 KB)
参考文献
[1]. H Stöppler, et al. The serine protease inhibitors TLCK and TPCK react with the RB-binding core of HPV-18 E7 protein and abolish its RB-binding capability. Virology. 1996 Mar 15;217(2):542-53. [Content Brief]
[2]. Patrícia de A Machado, et al. Effects of a Serine Protease Inhibitor N- p-Tosyl-L-phenylalanine Chloromethyl Ketone (TPCK) on Leishmania amazonensis and Leishmania infantum. Pharmaceutics.2022 Jun 29;14(7):1373. [Content Brief]
[3]. M H LeBlanc, et al. N-tosyl-L-phenylalanyl-chloromethylketone reduces hypoxic-ischemic brain injury in rat pups. Eur J Pharmacol. 2000 Mar 3;390(3):249-56 [Content Brief]
[4]. K Watanabe, et al. Vascular permeability changes by proteinase inhibitors in carrageenin-induced inflammation in rats. Agents Actions. 1986 Mar;17(5-6):472-7. [Content Brief]
Complete Stock Solution Preparation Table
Please refer to the solubility information to select the appropriate solvent. Once prepared, please aliquot and store the solution to prevent product inactivation from repeated freeze-thaw cycles.
Storage method and period of stock solution: -80°C, 6 months; -20°C, 1 month. When stored at -80°C, please use it within 6 months. When stored at -20°C, please use it within 1 month.
| Optional Solvent | Concentration Solvent Mass | 1 mg | 5 mg | 10 mg | 25 mg |
|---|---|---|---|---|---|
| DMSO | 1 mM | 2.8421 mL | 14.2106 mL | 28.4212 mL | 71.0530 mL |
| 5 mM | 0.5684 mL | 2.8421 mL | 5.6842 mL | 14.2106 mL | |
| 10 mM | 0.2842 mL | 1.4211 mL | 2.8421 mL | 7.1053 mL | |
| 15 mM | 0.1895 mL | 0.9474 mL | 1.8947 mL | 4.7369 mL | |
| 20 mM | 0.1421 mL | 0.7105 mL | 1.4211 mL | 3.5527 mL | |
| 25 mM | 0.1137 mL | 0.5684 mL | 1.1368 mL | 2.8421 mL | |
| 30 mM | 0.0947 mL | 0.4737 mL | 0.9474 mL | 2.3684 mL | |
| 40 mM | 0.0711 mL | 0.3553 mL | 0.7105 mL | 1.7763 mL | |
| 50 mM | 0.0568 mL | 0.2842 mL | 0.5684 mL | 1.4211 mL | |
| 60 mM | 0.0474 mL | 0.2368 mL | 0.4737 mL | 1.1842 mL | |
| 80 mM | 0.0355 mL | 0.1776 mL | 0.3553 mL | 0.8882 mL | |
| 100 mM | 0.0284 mL | 0.1421 mL | 0.2842 mL | 0.7105 mL |