Dansylcadaverine (Monodansyl cadaverine)
(Synonyms: 丹酰尸胺; Monodansyl cadaverine) 目录号 : GC30072
A fluorescent marker for autophagic vacuoles
Cas No.:10121-91-2
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
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Kinase experiment: |
To determine the time course of transglutamination, thymosin β4 (120 μM) is incubated with Dansylcadaverine (5 mM) in 70 μL buffer consisting of 10 mM Tris-HCl, pH 7.4, 15 mM CaCl2, 3 mM DTT. The reaction is started by addition of 0.1 U transglutaminase. Immediately after addition of the enzyme (t=0) and at indicated times,10 μL are taken from the mixture, diluted in 490 μL 0.1% TFA to stop the reaction and analyzed by HPLC[1]. |
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Cell experiment: |
MCF7 cells (2.4×104) are seeded into 35 mm plates. After 24 h incubation, CuO NPs are added with an increasing concentration in the presence or absence of 3-Methyladenine (3-MA) for different time periods . The cells are then incubated with 50 mM Dansylcadaverine (MDC) at 37°C for 15 min and washed with 1×PBS three times with 5 min interval. Finally, the cells are observed under a fluorescence microscope[2]. |
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References: [1]. Huff T, et al. Thymosin beta(4) serves as a glutaminyl substrate of transglutaminase. Labeling with fluorescentdansylcadaverine does not abolish interaction with G-actin. FEBS Lett. 1999 Dec 24;464(1-2):14-20. |
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Monodansylcadaverine (MDC) is a fluorescent marker for autophagic vacuoles.1,2 It is an autofluorescent substance incorporated into multilamellar bodies by both an ion trapping mechanism and interaction with membrane lipids, exhibiting a Stokes shift and increased relative fluorescence in hydrophobic environments.1 MDC selectively accumulates in PaTu 8902 subcellular fractions containing the lysosomal enzymes acid phosphatase and cathepsin D, but not those containing the rough and smooth endoplasmic reticulum markers TRAM and cytochrome P450, respectively.2
1.Niemann, A., Takatsuki, A., and Els?sser, H.P.The lysosomotropic agent monodansylcadaverine also acts as a solvent polarity probeJ. Histochem. Cytochem.48(2)251-258(2000) 2.Biederbick, A., Kern, H.F., and Els?sser, H.P.Monodansylcadaverine (MDC) is a specific in vivo marker for autophagic vacuolesEur. J. Cell Biol.66(1)3-14(1995)
| Cas No. | 10121-91-2 | SDF | |
| 别名 | 丹酰尸胺; Monodansyl cadaverine | ||
| Canonical SMILES | NCCCCCNS(=O)(C1=CC=CC2=C1C=CC=C2N(C)C)=O | ||
| 分子式 | C17H25N3O2S | 分子量 | 335.46 |
| 溶解度 | DMSO : 62.5 mg/mL (186.31 mM) | 储存条件 | Store at -20°C, protect from light, stored under nitrogen |
| 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 | 2.981 mL | 14.9049 mL | 29.8098 mL |
| 5 mM | 0.5962 mL | 2.981 mL | 5.962 mL |
| 10 mM | 0.2981 mL | 1.4905 mL | 2.981 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。
Effect of transglutaminase substrates and polyamines on the cellular sequestration and processing of follicle-stimulating hormone by rat Sertoli cells
Biol Reprod 1986 Aug;35(1):49-58.2874845 10.1095/biolreprod35.1.49
Transglutaminase (TGase) substrates Monodansyl cadaverine (MDC, monodansyl-1,5 diaminopentane) and methylamine (MA) and polyamines (PA) were tested for their effects on the cellular processing of radioiodinated human follicle-stimulating hormone (125I-hFSH). Specifically bound 125I-hFSH that could be released from cells during 10-min incubation period with acidified (pH 3.9) Hanks balanced-salt solution was considered membrane-bound unsequestered hormone. The rate at which cells sequestered 125I-hFSH into cellular compartments resistant to acid dissociation depended on the length of time in which cells were incubated with hormone. Cells incubated with 125I-hFSH for 15, 60, and 120 min had half-lives of sequestration of 26, 55 and 67 min respectively. One hundred-micromolar MDC inhibited degradation of 125I-hFSH as measured by the presence of radioactivity in the medium that was soluble in trichloroacetic acid. The rate of sequestration was never slower than that of controls, indicating that MDC did not decrease the ability of Sertoli cells to sequester 125I-hFSH. Despite these two observations, radioactivity associated with cells (acid-resistant radioactivity) was lower in cells treated with MDC than in controls. No effect of MDC on specific binding of 125I-hFSH was observed. Similar results were observed with MA, albeit at higher levels (0.0025-0.0425 M), consistent with their relative potency to inhibit TGase activity. Polyamines, spermine, and putrescine also decreased cell-associated radioactivity despite decreasing degradation of hFSH. TGase substrates (MDC, MA, PA) prevented entry of sequestered 125I-hFSH into the degradative pathways of Sertoli cells. These data suggest that transglutamination may influence the fate of sequestered FSH in Sertoli cells but not the rate at which sequestration occurs.
Inhibition of receptor internalization attenuates the TNFalpha-induced ROS generation in non-phagocytic cells
Biochem Biophys Res Commun 2006 Dec 29;351(4):972-8.17097052 10.1016/j.bbrc.2006.10.154
Reactive oxygen species (ROS) are important regulatory molecules implicated in the signaling cascade triggered by tumor necrosis factor (TNF)alpha, although the events through which TNFalpha induces ROS generation are not well characterized. Here, we report that TNFalpha-induced ROS production was blocked by pretreatment with internalization inhibitor Monodansyl cadaverine (MDC). Similarly, a transient expression of a GTP-binding and hydrolysis-defective dynamin mutant (dynamin(K44A)) that had been shown to be defective in internalization significantly attenuated the TNFalpha-induced intracellular ROS production. Importantly, the inhibition of receptor internalization suppressed TNFalpha signaling to mitogen-activated protein kinases (MAPKs) stimulation. Together, our results suggest that receptor internalization is somehow necessary for the TNFalpha-induced ROS generation and subsequent intracellular downstream signaling in non-phagocytes.
Cross linking to tissue transglutaminase and collagen favours gliadin toxicity in coeliac disease
Gut 2006 Apr;55(4):478-84.16188922 PMC1856150
Background and aims: Intestinal inflammation in coeliac disease is driven by the gluten fraction of wheat proteins. Deamidation or cross linking of gluten peptides by tissue transglutaminase (tTG), the coeliac disease autoantigen, creates potent T cell stimulatory peptides. Therefore, our aim was to identify the reaction patterns of gluten peptides, intestinal extracellular matrix proteins, and tTG. Methods: tTG activity was analysed by incorporation of Monodansyl cadaverine into gliadins. Fluorescence labelled tTG reactive short gliadin peptides were used to demonstrate their deamidation and explore their cross linking patterns with tTG itself or extracellular matrix proteins. Patient sera and controls were checked for autoantibodies to matrix proteins. Results: Gliadins alpha1-alpha11, gamma1-gamma6, omega1-omega3, and omega5 were substrates for tTG. tTG catalysed the cross linking of gliadin peptides with interstitial collagen types I, III, and VI. Coeliac patients showed increased antibody titres against the collagens I, III, V, and VI. Conclusions: tTG formed high molecular weight complexes with all tested gliadins. As all tested gliadins were substrates for tTG, the tTG catalysed modifications were not restricted to single gliadin types and epitopes. Furthermore, haptenisation and long term immobilisation of gliadin peptides by tTG catalysed binding to abundant extracellular matrix proteins could be instrumental in the perpetuation of intestinal inflammation and some associated autoimmune diseases in coeliac disease.
Suppression of aggregate formation and apoptosis by transglutaminase inhibitors in cells expressing truncated DRPLA protein with an expanded polyglutamine stretch
Nat Genet 1998 Feb;18(2):111-7.9462738 10.1038/ng0298-111
To elucidate the molecular mechanisms whereby expanded polyglutamine stretches elicit a gain of toxic function, we expressed full-length and truncated DRPLA (dentatorubral-pallidoluysian atrophy) cDNAs with or without expanded CAG repeats in COS-7 cells. We found that truncated DRPLA proteins containing an expanded polyglutamine stretch form filamentous peri- and intranuclear aggregates and undergo apoptosis. The apoptotic cell death was partially suppressed by the transglutaminase inhibitors cystamine and Monodansyl cadaverine (but not putrescine), suggesting involvement of a transglutaminase reaction and providing a potential basis for the development of therapeutic measures for CAG-repeat expansion diseases.
Receptor-mediated endocytosis of IL-8: a fluorescent microscopic evidence and implication of the process in ligand-induced biological response in human neutrophils
Cytokine 1997 Aug;9(8):587-96.9245487 10.1006/cyto.1997.0206
Interleukin 8 (IL-8), a neurophil-activating and chemotactic cytokine, is known to play a key role in the pathogenesis of a large number of neutrophil-driven inflammatory diseases. Although the cytokine is rapidly internalized at 37 degrees C with its receptors, there was no direct evidence for the ligand-induced endocytosis of the receptor or that of the interaction of receptor ligand complex at 37 degrees C. As a result, our understanding about the regulation of Il-8 induced biological response is very limited. In the present study, using FITC-IL-8 conjugate as a probe, we have demonstrated the time- and temperature-dependent endocytosis of IL-8 under fluorescent microscope. We have also shown that the bright fluorescent light on the surface of neutrophils gradually disappears and it becomes almost dark after 120 min of incubation. Monodansyl cadaverine (MDC, 900 microM), however, was found to retain the fluorescent light of FITC coupled with Il-8 on the cells. MDC and ouabain (2.5 mM) can inhibit the ligand induced endocytosis by 76% and 96%, respectively, compared to control. With respect to control, IL-8 induced biological responses e.g. IL-8 directed migration, intracellular Ca2+ release and superoxide release are significantly reduced by 77%, 94% and 76%, respectively, in presence of MDC. The study presents a direct visual evidence of the time and temperature-dependent receptor-mediated endocytosis of IL-8 which is inhibited by MDC and ouabain. This information is useful for understanding the ligand receptor interaction at 37 degrees C and may be useful for developing anti-inflammatory agents against IL-8.