Tosyl Phenylalanyl Chloromethyl Ketone
(Synonyms: L-1,4'-甲基磺酰基-2-苯基乙基氯甲基酮,L-1-Tosylamido-2-phenylethyl chloromethyl ketone; L-TPCK) 目录号 : GC45066
An inhibitor of chymotrypsin-like serine proteases
Cas No.:402-71-1
Sample solution is provided at 25 µL, 10mM.
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Quality Control & SDS
- View current batch:
- Purity: >97.00%
- COA (Certificate Of Analysis)
- SDS (Safety Data Sheet)
- Datasheet
Tosyl phenylalanyl chloromethyl ketone (TPCK) is an irreversible inhibitor of chymotrypsin-like proteases that has been shown to affect cell proliferation, apoptosis, and tumorigenesis. It can disrupt PDK1 signaling to the AGC kinases, Akt, S6K1, and RSK, as well as MSK1 and MSK2. TPCK also inhibits superoxide production and suppresses neutrophil respiratory burst.
| Cas No. | 402-71-1 | SDF | |
| 别名 | L-1,4'-甲基磺酰基-2-苯基乙基氯甲基酮,L-1-Tosylamido-2-phenylethyl chloromethyl ketone; L-TPCK | ||
| Canonical SMILES | CC1=CC=C(S(N[C@H](C(CCl)=O)CC2=CC=CC=C2)(=O)=O)C=C1 | ||
| 分子式 | C17H18ClNO3S | 分子量 | 351.8 |
| 溶解度 | DMF: 30 mg/ml,DMSO: 30 mg/ml,DMSO:PBS(pH 7.2) (1:3): 0.25 mg/ml,Ethanol: 10 mg/ml | 储存条件 | Store at -20°C |
| General tips | 请根据产品在不同溶剂中的溶解度选择合适的溶剂配制储备液；一旦配成溶液，请分装保存，避免反复冻融造成的产品失效。 储备液的保存方式和期限：-80°C 储存时，请在 6 个月内使用，-20°C 储存时，请在 1 个月内使用。 为了提高溶解度，请将管子加热至37℃，然后在超声波浴中震荡一段时间。 |
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| Shipping Condition | 评估样品解决方案：配备蓝冰进行发货。所有其他可用尺寸：配备RT，或根据请求配备蓝冰。 | ||
| 制备储备液 | |||
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1 mg | 5 mg | 10 mg |
| 1 mM | 2.8425 mL | 14.2126 mL | 28.4252 mL |
| 5 mM | 0.5685 mL | 2.8425 mL | 5.685 mL |
| 10 mM | 0.2843 mL | 1.4213 mL | 2.8425 mL |
| 第一步：请输入基本实验信息（考虑到实验过程中的损耗，建议多配一只动物的药量） | ||||||||||
| 给药剂量 | mg/kg |  |
动物平均体重 | g | |
每只动物给药体积 | ul | |
动物数量 | 只 |
| 第二步：请输入动物体内配方组成（配方适用于不溶于水的药物；不同批次药物配方比例不同，请联系GLPBIO为您提供正确的澄清溶液配方） | ||||||||||
| % DMSO % % Tween 80 % saline | ||||||||||
| 计算重置 | ||||||||||
计算结果:
工作液浓度： mg/ml；
DMSO母液配制方法： mg 药物溶于 μL DMSO溶液（母液浓度 mg/mL，
体内配方配制方法：取 μL DMSO母液，加入 μL PEG300，混匀澄清后加入μL Tween 80，混匀澄清后加入 μL saline，混匀澄清。
1. 首先保证母液是澄清的；
2.
一定要按照顺序依次将溶剂加入，进行下一步操作之前必须保证上一步操作得到的是澄清的溶液，可采用涡旋、超声或水浴加热等物理方法助溶。
3. 以上所有助溶剂都可在 GlpBio 网站选购。
Ornithine decarboxylase inactivation in HeLa cells
J Cell Physiol 1976 Sep;89(1):65-76.PMID:956282DOI:10.1002/jcp.1040890107.
Ornithine decarboxylase (ODC) can be induced up to 100-fold over basal levels four hours after addition of glutamine to the medium of HeLa cells growing in suspension culture. As demonstrated in several other cell types, ODC is inactivated very rapidly in HeLa cells, and the rate of inactivation is seen to vary with a half life of 9-15 minutes in uninduced cells and rises to ca. 60 minutes at the peak of induction. Quantitatively, the change in rate of inactivation cannot completely account for the observed rise in activity, thus synthesis or activation of ODC must also be involved in the induction process. The inactivation process requires metabolic energy and it can be sustained by glycolytic derived energy. Other factors which are known to inhibit protein breakdown in mammalian cells, such as sodium fluoride, insulin, or Tosyl Phenylalanyl Chloromethyl Ketone, had no effect on the rate of inactivation of ODC. Attempts to demonstrate ODC inactivation in a cell free system at neutral pH were unsuccessful.
Genome-Wide Mutant Screening in Yeast Reveals that the Cell Wall is a First Shield to Discriminate Light From Heavy Lanthanides
Front Microbiol 2022 May 19;13:881535.PMID:35663896DOI:10.3389/fmicb.2022.881535.
The rapidly expanding utilization of lanthanides (Ln) for the development of new technologies, green energies, and agriculture has raised concerns regarding their impacts on the environment and human health. The absence of characterization of the underlying cellular and molecular mechanisms regarding their toxicity is a caveat in the apprehension of their environmental impacts. We performed genomic phenotyping and molecular physiology analyses of Saccharomyces cerevisiae mutants exposed to La and Yb to uncover genes and pathways affecting Ln resistance and toxicity. Ln responses strongly differed from well-known transition metal and from common responses mediated by oxidative compounds. Shared response pathways to La and Yb exposure were associated to lipid metabolism, ion homeostasis, vesicular trafficking, and endocytosis, which represents a putative way of entry for Ln. Cell wall organization and related signaling pathways allowed for the discrimination of light and heavy Ln. Mutants in cell wall integrity-related proteins (e.g., Kre1p, Kre6p) or in the activation of secretory pathway and cell wall proteins (e.g., Kex2p, Kex1p) were resistant to Yb but sensitive to La. Exposure of WT yeast to the serine protease inhibitor Tosyl Phenylalanyl Chloromethyl Ketone mimicked the phenotype of kex2∆ under Ln, strengthening these results. Our data also suggest that the relative proportions of chitin and phosphomannan could modulate the proportion of functional groups (phosphates and carboxylates) to which La and Yb could differentially bind. Moreover, we showed that kex2∆, kex1∆, kre1∆, and kre6∆ strains were all sensitive to light Ln (La to Eu), while being increasingly resistant to heavier Ln. Finally, shotgun proteomic analyses identified modulated proteins in kex2∆ exposed to Ln, among which several plasmalemma ion transporters that were less abundant and that could play a role in Yb uptake. By combining these different approaches, we unraveled that cell wall components not only act in Ln adsorption but are also active signal effectors allowing cells to differentiate light and heavy Ln. This work paves the way for future investigations to the better understanding of Ln toxicity in higher eukaryotes.
Establishment of a rapid ELISPOT assay for influenza virus titration and neutralizing antibody detection
J Med Virol 2021 Jun;93(6):3455-3464.PMID:32621615DOI:10.1002/jmv.26257.
Seasonal influenza is an acute respiratory infection causing around 500 000 global deaths annually. There is an unmet medical need to develop more effective antiviral drugs and vaccines against influenza infection. A rapid, accurate, high-throughput titration assay for influenza virus particles or neutralizing antibodies would be extremely useful in these research fields. However, commonly used methods such as tissue culture infective dose and plaque-forming units (PFU) for virus particle quantification, and the plaque reduction neutralization test (PRNT) for antibody determination are time-consuming, laborious, and have limited accuracy. In this study, we developed an efficient assay based on the enzyme-linked immunospot (ELISPOT) technique for the influenza virus and neutralizing antibody titration. Two broad-spectrum antibodies recognizing the nucleoproteins of influenza A and B viruses were used in the assay to broadly and highly sensitively detect influenza virus-infected cells at 16 hours postinfection. An optimized cell culture medium with no Tosyl Phenylalanyl Chloromethyl Ketone trypsin and high dose oseltamivir acid was used to improve quantitation accuracy. This ELISPOT assay displayed a good correlation (R2 = 0.9851) with the PFU assay when used to titrate 30 influenza virus isolates. The assay was also applied to measure influenza-neutralizing antibodies in 40 human sera samples, showing a good correlation (R2 = 0.9965) with the PRNT assay. This ELISPOT titration assay is a rapid, accurate, high-throughput assay for quantification of influenza virus and neutralizing antibodies, and provides a powerful tool for research into and development of drugs and vaccines against influenza.
Epinephrine evokes renalase secretion via α-adrenoceptor/NF-κB pathways in renal proximal tubular epithelial cells
Kidney Blood Press Res 2014;39(4):252-9.PMID:25171187DOI:10.1159/000355802.
Background/aims: Renalase is a recently discovered, kidney-specific monoamine oxidase that metabolizes circulating catecholamines. These findings present new insights into hypertension and chronic kidney diseases. Previous data demonstrated that renalase was mainly secreted from proximal tubules which could be evoked by catecholamines. The purpose of this study is to investigate whether renalase expression is induced by epinephrine via α-adrenoceptor/NFκB pathways. Methods: HK2 cells were utilized to explore renalase expression in response to epinephrine in vitro. Phentolamine, an α-adrenoceptor antagonist, and Tosyl Phenylalanyl Chloromethyl Ketone (TPCK) were used to block α-adrenoceptor and to knock down the transcription factor NFκB, respectively. Renalase expression was analyzed using Western blot and quantitative PCR. Results: Both protein and mRNA levels of renalase in HK2 cells increased in response to epinephrine (P<0.05). Epinephrine-evoked renalase expression was attenuated by phentolamine and TPCK separately (P<0.05). Conclusion: Epinephrine evokes renalase secretion via α-adrenoceptor/NF-κB pathways in renal proximal tubular epithelial cells.
Generation and identification of anti-inflammatory peptides from bovine β-casein using enzyme preparations from cod and hog
J Sci Food Agric 2016 Feb;96(3):868-77.PMID:25754585DOI:10.1002/jsfa.7159.
Background: The aim of the present study was to generate and identify potential anti-inflammatory peptides from bovine β-casein with enzyme preparations from cod and hog. Furthermore, the potential of cod trypsin, derived from fishery by-products, to produce these bioactive peptides for replacement of non-food-grade Tosyl Phenylalanyl Chloromethyl Ketone (TPCK)-treated porcine trypsin enzyme preparation was evaluated. Results: Potential anti-inflammatory peptides were obtained by hydrolysis of β-casein with the tryptic enzyme preparations cod trypsin, porcine trypsin (TPCK-treated) and a porcine trypsin and chymotrypsin preparation (PTN 6.0 S). Proteolysates generated with enzyme preparations containing mainly chymotryptic activity (Cryotin, Cryotin F) did not exhibit any effect. Conclusion: The more chymotryptic enzyme activity is present, the lower is the potential anti-inflammatory activity of the hydrolysates in HEK(nfκb-RE) cells. Comparable peptides were produced by application of porcine trypsin (TPCK) and cod trypsin. Therefore, the enzyme preparation cod trypsin can replace the non-food-grade porcine enzyme preparation trypsin (TPCK) for the generation of potential anti-inflammatory peptides from β-casein.