DiD perchlorate
(Synonyms: 1,1'-双十八烷基-3,3,3',3'-四甲基吲哚二碳菁高氯酸盐) 目录号 : GC30543DiD perchlorate is a lipophilic fluorescent dye that can rapidly and stably integrate into phospholipid cell membranes and is widely used as Di to label cells, organelles, liposomes, viruses and lipoproteins
Cas No.:127274-91-3
Sample solution is provided at 25 µL, 10mM.
Quality Control & SDS
- View current batch:
- Purity: >95.00%
- COA (Certificate Of Analysis)
- SDS (Safety Data Sheet)
- Datasheet
本方案仅提供一个指导,应根据您的具体需要进行修改。
1.细胞膜染色液制备
(1)配置DMSO或EtOH 储存液:储存液用DMSO或EtOH配置,浓度1~5mM。例如,取25mg DiD(Mw:959.91g/mol)溶于5.21ml无水DMSO中,充分溶解,即得到5mM的储存液。
注意:未使用的储存液分装保存在-20℃,避免反复冻融。
(2)工作液制备:用合适的缓冲液(如:无血清培养基,HBSS或PBS)稀释储存液,配制浓度为0.5~5μM的工作液。
注意: 工作液终浓度需要根据不同细胞系和实验体系来优化,建议从推荐浓度开始,以10倍范围为区间进行最优浓度的摸索。
2.悬浮细胞染色
(1)悬浮细胞经4°C、1000-1500rpm离心3-5分钟,弃去上清液。用PBS清洗两次,每次5分钟。
(2)加入1mL的DiD perchlorate工作溶液,室温孵育5-30分钟。
注意:不同的细胞最佳培养时间不同,可以20min作为起始孵育时间,之后优化以保证得到均一化的标记结果。
(3)孵育结束后,经1000-1500rpm离心5分钟,去除上清液,加入PBS清洗2-3次,每次5分钟。
(4)用预温的无血清细胞培养基或PBS重悬细胞。通过荧光显微镜或流式细胞术观察。
3.粘壁细胞染色
(1)在无菌盖玻片上培养贴壁细胞。
(2)从培养基中移走盖玻片,吸出过量的培养基,将盖玻片放在潮湿的环境中。
(3)从盖玻片的一角加入100uL的染料工作液,轻轻晃动使染料均匀覆盖所有细胞。
(4)室温条件下孵育5-30分钟。不同的细胞最佳培养时间不同,可以20min作为起始孵育时间,之后优化以保证得到均一化的标记结果。
(5)孵育结束后吸弃染料工作液,用预温的培养液洗盖玻片2~3次。
4.显微镜检测:DiD perchlorate的激发/发射光分别为650/670nm。
注意事项:
1)荧光染料均存在淬灭问题,请尽量注意避光,以减缓荧光淬灭。
2)为了您的安全和健康,请穿实验服并戴一次性手套操作。
DiD perchlorate is a lipophilic fluorescent dye that can rapidly and stably integrate into phospholipid cell membranes and is widely used as Di to label cells, organelles, liposomes, viruses and lipoproteins[1]. DiD exhibits distinct red fluorescence, which facilitates multicolor imaging and flow cytometry analysis of live cells[2]. DiD perchlorate has been used to label plasma membranes and endocytic organelles in bovine aortic endothelial cells and rat hippocampal slices[3]. DiD has also been used to assess proliferation in prostate cancer cell lines by flow cytometry, where cell populations with high DiD expression were associated with lower proliferation[3]. DiD is non-cytotoxic and detectable after three weeks in subcutaneously implanted PC3 cells in vivo[4]. DiD can be excited by 633nm helium-neon (He-Ne) laser, has a longer excitation and emission wavelength than Dil, and is especially suitable for labeling cells and tissues with background fluorescence.
DiD高氯酸盐是一种亲脂性荧光染料,它可以快速稳定地整合到磷脂细胞膜中,被广泛用作Di来标记细胞,细胞器,脂质体,病毒和脂蛋白[1]。DiD表现出明显的红色荧光,有利于活细胞的多色成像和流式细胞术分析[2]。DiD高氯酸盐已被用于标记牛主动脉内皮细胞和大鼠海马切片中的质膜和内吞细胞器[3]。DiD还被用于通过流式细胞术评估前列腺癌细胞系中的增殖,其中高DiD表达的细胞群与较低的增殖有关[3]。DiD无细胞毒性,可在三周后在体内皮下植入的PC3细胞中检测到[4]。DiD可被633nm氦-氖(He-Ne)激光激发,具有比Dil更长的激发和发射波长,特别适合用于标记具本底荧光的细胞和组织。
References:
[1]. Yumoto, K., Berry, J.E., Taichman, R.S., et al.A novel method for monitoring tumor proliferation in vivo using fluorescent dye DiDCytometry A.85(6)548-555(2014).
[2]. Dailey, M.E., and Waite, M.Confocal imaging of microglial cell dynamics in hippocampal slice culturesMethods18(2)222-230(1999).
[3]. Lin, C.P., Lynch, M.C., and Kochevar, I.E.Reactive oxidizing species produced near the plasma membrane induce apoptosis in bovine aorta endothelial cellsExp. Cell Res.259(2)351-359(2000).
[4]. Ribeiro, T., Raja, S., Rodrigues, A.S., et al.NIR and visible perylenediimide-silica nanoparticles for laser scanning bioimagingDyes Pigments110227-234(2014)
Cas No. | 127274-91-3 | SDF | |
别名 | 1,1'-双十八烷基-3,3,3',3'-四甲基吲哚二碳菁高氯酸盐 | ||
Canonical SMILES | CCCCCCCCCCCCCCCCCCN1C2=C(C=CC=C2)C(C)(C)/C1=C\C=C\C=C\C3=[N+](C4=C(C=CC=C4)C3(C)C)CCCCCCCCCCCCCCCCCC.O=Cl(=O)([O-])=O | ||
分子式 | C61H99ClN2O4 | 分子量 | 959.9 |
溶解度 | DMSO: 50 mg/mL (52.09 mM) | 储存条件 | Store at -20°C,protect from light, stored under nitrogen,unstable in solution, ready to use. |
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 | |||
1 mg | 5 mg | 10 mg | |
1 mM | 1.0418 mL | 5.2089 mL | 10.4178 mL |
5 mM | 0.2084 mL | 1.0418 mL | 2.0836 mL |
10 mM | 0.1042 mL | 0.5209 mL | 1.0418 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。
Perchlorate in Water Supplies: Sources, Exposures, and Health Effects
Perchlorate exposure occurs from ingestion of natural or man-made perchlorate in food or water. Perchlorate is used in a variety of industrial products including missile fuel, fireworks, and fertilizers, and industrial contamination of drinking water supplies has occurred in a number of areas. Perchlorate blocks iodide uptake into the thyroid and decreases the production of thyroid hormone, a critical hormone for metabolism, neurodevelopment, and other physiologic functions. Occupational and clinical dosing studies have not identified clear adverse effects, but may be limited by small sample sizes, short study durations, and the inclusion of mostly healthy adults. Expanding evidence suggests that young children, pregnant women, fetuses, and people co-exposed to similarly acting agents may be especially susceptible to perchlorate. Given the ubiquitous nature of perchlorate exposure, and the importance of thyroid hormone for brain development, studying the impact of perchlorate on human health could have far-reaching public health implications.
Perchlorate and Diet: Human Exposures, Risks, and Mitigation Strategies
Perchlorate is an endocrine-disrupting chemical that interferes with the normal functioning of the thyroid gland. Maternal thyroid dysfunction during gestation may alter fetal brain development. Perchlorate contamination is widespread: it is present in the body of all Americans tested and the majority of foods tested. The main sources of food contamination appear to be hypochlorite bleach, a disinfectant and sanitizer, that when poorly managed quickly degrades to perchlorate and perchlorate-laden plastic food packaging for dry food or localized contamination from manufacturing or processing of the chemical. Eliminating perchlorate from food packaging and improving bleach management, such as reducing concentration and storage time and temperature, would result in reduced perchlorate contamination of food and water.
Perchlorate as an emerging contaminant in soil, water and food
Perchlorate ( [Formula: see text] ) is a strong oxidizer and has gained significant attention due to its reactivity, occurrence, and persistence in surface water, groundwater, soil and food. Stable isotope techniques (i.e., ((18)O/(16)O and (17)O/(16)O) and (37)Cl/(35)Cl) facilitate the differentiation of naturally occurring perchlorate from anthropogenic perchlorate. At high enough concentrations, perchlorate can inhibit proper function of the thyroid gland. Dietary reference dose (RfD) for perchlorate exposure from both food and water is set at 0.7 μg kg(-1) body weight/day which translates to a drinking water level of 24.5 μg L(-1). Chromatographic techniques (i.e., ion chromatography and liquid chromatography mass spectrometry) can be successfully used to detect trace level of perchlorate in environmental samples. Perchlorate can be effectively removed by wide variety of remediation techniques such as bio-reduction, chemical reduction, adsorption, membrane filtration, ion exchange and electro-reduction. Bio-reduction is appropriate for large scale treatment plants whereas ion exchange is suitable for removing trace level of perchlorate in aqueous medium. The environmental occurrence of perchlorate, toxicity, analytical techniques, removal technologies are presented.
Perchlorate removal by a combined heterotrophic and bio-electrochemical hydrogen autotrophic system
Here, a novel combined heterotrophic and bio-electrochemical hydrogen autotrophic (CHBHA) system was developed to remove perchlorate under low chemical dosages and energy consumption. The perchlorate removal performance at various hydraulic retention times (HRTs) and acetate dosages was investigated. For influent containing 10 ± 0.10 mg/L perchlorate, the optimal removal efficiency by the CHBHA system was 98.96 ± 1.62 %, 92.99 ± 2.99 %, 97.85 ± 0.41 %, and 98.24 ± 1.56 % at different operating stages. Perchlorate was mainly removed in the heterotrophic part (H-part) at a sufficient HRT (6 h) and acetate dosage (14.75 mg/L). At other stages, perchlorate was synergistically removed by the H-part and electrochemical hydrogen autotrophic part (E-part). Since the H-part removed some perchlorate, the E-part's applied current decreased, thus reducing energy costs. The maximum current efficiency of CHBHA system was 22.09 %. Compared with the single E-part system, the combined system used 65 % less energy. Perchlorate was converted into active chlorine in the E-part, which improved the effluent quality. The bacterial community structures of the two parts were significantly different. Comamonas, Dechloromonas, Acinetobacter, and Chryseobacterium were enriched in the H-part, and the dominant genera in the E-part were Thauera, Azonexus, Hydrogenophaga, and Tissierella.
Potassium perchlorate effects on primordial germ cells of developing medaka larvae
Perchlorate is a chemical compound commonly used in military artillery and equipment. It has been detected in drinking water, air, soil, and breast milk. Exposure of humans to perchlorate can occur in the theater of war and areas adjacent to military training grounds. A high concentration of perchlorate has been found to affect reproduction in vertebrates, including fish. However, whether environmental concentrations of perchlorate can affect primordial germ cells (PGCs), the founders of sperm and eggs, is not clearly understood. In the present study, we examined the effects of 0, 10, 100, and 1000 μg/L potassium perchlorate exposure on the embryonic development of medaka and their PGCs. Perchlorate exposure delayed hatching time, reduced heartbeat, inhibited migration of PGCs, and increased developmental deformities in the larvae. The 10 and 20 mg/L concentrations of perchlorate were lethal to embryos, whereas vitamin C co-treatment (1 mg/L) completely blocked perchlorate-induced mortality. RNA-seq analysis of isolated PGCs showed a non-linear pattern in expression profiles of differentially altered genes. Significantly upregulated genes were found in PGCs from the 10 and 1000 μg/L groups, whereas the 100 μg/L groups showed the highest number of significantly downregulated genes. Gene ontology analysis predicted differentially expressed genes to be involved in proteolysis, metabolic processes, peptides activity, hydrolase activity, and hormone activity. Among the cellular components, extracellular, intracellular, sarcoplasmic, and 6-phosphofructokinase and membrane-bounded processes were affected. Ingenuity Pathway Analysis of PGC transcriptomes revealed thyroid hormone signaling to be affected by all concentrations of perchlorate. The present results suggested that perchlorate affected the development of medaka larvae and vitamin C was able to ameliorate perchlorate-induced embryo mortality. Additionally, perchlorate altered the global transcriptional network in PGCs in a non-linear fashion suggesting its potential effects on developing germ cells and fertility.