Tosyllysine Chloromethyl Ketone (hydrochloride)
(Synonyms: (3S)-7-氨基-1-氯-3-磺酰氨基-2-庚酮盐酸盐,1-Chloro-3-Toxylamide-7-Amino-L-2-Heptanone,TLCK) 目录号 : GC13967A non-selective proteinase inhibitor
Cas No.:4272-74-6
Sample solution is provided at 25 µL, 10mM.
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Tosyllysine Chloromethyl Ketone (hydrochloride) is a protease inhibitor [1][2][3].
L-1-chloro-3-[4-tosylamido]-7-amino-2-heptanone-HCl (TLCK) is a protease inhibitor. TLCK is an active site-directed agent that inhibits serine proteinases with trypsin-like activity. TLCK also interacts non-selectively with thiol groups and thereby inhibits cysteine proteinases and other enzymes. In LPS-activated rat alveolar macrophages, TLCK at 1-100 μM inhibited NOx- accumulation and inducible iNOS expression in a concentration-dependent way [1]. To prevent proteolytic degradation, TLCK may be used in protein purification protocols [2]. TLCK significantly increased the cytotoxic activity of C. histolyticum supernatant towards human epithelial HeLa cells probably by hindering natural defence mechanisms of cells. 30 min incubation with bacterial supernatant increased toxicity in both concentrations (200 and 1000 μM) from 18 ± 3% to 39 ± 3% and 57 ± 8%, respectively. TLCK also blocked clostripain enzymatic activity obtained from C. histolyticum. So TLCK might be used to treat diseases complicated by concurrent C. histolyticum infection [3].
References:
[1]. Griscavage JM, Wilk S, Ignarro LJ. Serine and cysteine proteinase inhibitors prevent nitric oxide production by activated macrophages by interfering with transcription of the inducible NO synthase gene. Biochem Biophys Res Commun. 1995 Oct 13;215(2):721-9.
[2]. Urban MK, Franklin SG, Zweidler A. Isolation and characterization of the histone variants in chicken erythrocytes. Biochemistry. 1979 Sep 4;18(18):3952-60.
[3]. Józwiak, J.,Komar, A.,Jankowska, E., et al. Determination of the cytotoxic effect of Clostridium
histolyticum culture supernatant on HeLa cells in the presence of protease inhibitors. FEMS Immunology & Medical Microbiology 45(2):137-42 (2005).
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Cell experiment: |
HeLa cell line (human cervical cancer cells) is cultured in DMEM supplemented with 10% fetal bovine serum (FBS), L-glutamine (300 mg/L), penicillin (100 U/ml) and streptomycin(100 μg/ml) at 37°C in 5% CO2. HT-29 cell line (human colorectal adenocarcinoma) is cultured in RPMI supplemented with 10% fetal bovine serum (FBS), L-glutamine (300 mg/L), penicillin (100 U/mL) and streptomycin (100 μg/mL) at 37°C in 5% CO2. The cells are split every second day to keep the cell growth in logarithmic phase. The cells are routinely tested for mycoplasma. The cells are treated with different concentrations of TPCK or TLCK (5, 10, 25, 50, 100, 150, and 200 μM) for 30 min and/or with different concentrations of IFN-γ for 2 h followed by treatment with different concentrations of anti-Fas for 2 or 4 days for each specific experiment. The control cells are treated with the respective vehicle only. Cell viability analysis of HeLa and HT-29 cells is assessed by their XTT reduction activity. 100 µL of 2×104 cells/mL is incubated with treatments at the indicated time. At the end of the incubation period, 25 µL of 1 mg/mL XTT solution (containing 0.2 mM phenazine methosulphate (PMS) is added and the cells are incubated for an additional 1 h. The OD values are measured using an ELISA reader at 450 nm with a reference wavelength of 650 nm[1]. |
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References: [1]. Shadrin N, et al. Serine protease inhibitors interact with IFN-γ through up-regulation of FasR; a novel therapeutic strategy against cancer. Exp Cell Res. 2015 Jan 15;330(2):233-9. |
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| Cas No. | 4272-74-6 | SDF | |
| 别名 | (3S)-7-氨基-1-氯-3-磺酰氨基-2-庚酮盐酸盐,1-Chloro-3-Toxylamide-7-Amino-L-2-Heptanone,TLCK | ||
| 化学名 | N-[5-amino-1-(2-chloroacetyl)pentyl]-4-methyl-benzenesulfonamide, monohydrochloride | ||
| Canonical SMILES | CC1=CC=C(S(N[C@@H](CCCCN)C(CCl)=O)(=O)=O)C=C1.Cl | ||
| 分子式 | C14H21ClN2O3S • HCl | 分子量 | 369.3 |
| 溶解度 | ≥ 16.3mg/mL in Water | 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 | 2.7078 mL | 13.5391 mL | 27.0783 mL |
| 5 mM | 0.5416 mL | 2.7078 mL | 5.4157 mL |
| 10 mM | 0.2708 mL | 1.3539 mL | 2.7078 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。
Quality Control & SDS
- View current batch:
- Purity: >98.00%
- COA (Certificate Of Analysis)
- SDS (Safety Data Sheet)
- Datasheet