3,3'-Dihexyloxacarbocyanine iodide (DiOC6(3) iodide)
(Synonyms: 碘代3,3'-二己氧基羰花青,DiOC6(3) iodide) 目录号 : GC33493
A fluorescent dye
Cas No.:53213-82-4
Sample solution is provided at 25 µL, 10mM.
;)
,-1-7$$$37316672.jpg');)
;)
;)
$$$36806353.jpg');)
;33(7)%EF%BC%9A546-561$$$37156877.jpg');)
;)
;)
;)
%201124-1138.$$$35908550.jpg');)
%20766-780.$$$37100057.jpg');)
%20418-432.$$$36893736.jpg');)
%EF%BC%9A-240-255.$$$35108512.jpg');)
533-545$$$36543525.jpg');)
%201-13.$$$36600053.jpg');)
;)
Quality Control & SDS
- View current batch:
- Purity: >98.00%
- COA (Certificate Of Analysis)
- SDS (Safety Data Sheet)
- Datasheet
本方案仅提供一个指导，请根据您的具体需要进行修改。
1.制备染色液
(1)配置DMSO储存液:使用DMSO溶解DiOC6(3) iodide，配置浓度为1~5mM的储存液。
注意:未使用的储存液分装后在-20℃避光保存，避免反复冻融。
(2)配置工作液:用合适的缓冲液(如:无血清培养基或PBS)稀释储存液，配制浓度为0.5~5μM的工作液。
注意:请根据实际情况调整工作液浓度，现用现配。
2.细胞悬浮染色
(1)悬浮细胞：经4°C、1000g离心3-5分钟，弃去上清液，用PBS清洗两次，每次5分钟。
(2)贴壁细胞：使用PBS清洗两次，加入胰酶消化细胞，消化完成后经1000g离心3-5min。
(3)加入1mL的DiOC6(3) iodide工作溶液重悬约106个细胞，室温避光孵育5-30分钟。不同细胞最佳孵育时间不同，请根据具体实验需求自行摸索。
(4)孵育结束后，经1000g离心5分钟，去除上清液，加入PBS清洗2-3次，每次5分钟。
(5)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.细胞贴壁染色
(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100μL的染料工作液，轻轻晃动使染料均匀覆盖所有细胞。
(4)室温避光孵育5-30分钟。不同细胞最佳孵育时间不同，请根据具体实验需求自行摸索。
(5)孵育结束后吸弃染料工作液，使用预温的培养液清洗盖玻片2~3次。
4.显微镜检测：DiOC6(3) iodide的最大激发光/发射光分别为484/501 nm。
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。
3,3'-Dihexyloxacarbocyanine (DiOC6(3)) is a lipophilic fluorescent dye with excitation/emission spectra of 484/501 nm, respectively.1 At high concentrations, DiOC6(3) accumulates in the endoplasmic reticulum (ER) and has been used to visualize ER in moss, yeast, and muscle cells. At low concentrations, DiOC6(3) accumulates in mitochondria and has been used to assess mitochondrial dislocations, fusion, and fission in living cells, as well as to visualize cellular apoptosis. DiOC6(3) can also be used to assess cell membrane potential, as depolarization directly correlates with increased fluorescence.2,3 DiOC6(3) photoexcitation inactivates intracellular organelle movement and inhibits microtubule polymerization in vitro.4
1.Sabnis, R.W., Deligeorgiev, T.G., Jachak, M.N., et al.DiOC6(3): A useful dye for staining the endoplasmic reticulumBiotech. Histochem.72(5)253-258(1997) 2.Jenssen, H.-L., Redmann, K., and Mix, E.Flow cytometric estimation of transmembrane potential of macrophages--a comparison with microelectrode measurementsCytometry7(4)339-346(1986) 3.Hoffman, J.F., and Laris, P.C.Determination of membrane potentials in human and Amphiuma red blood cells by means of fluorescent probeJ. Physiol.239(3)519-552(1974) 4.Lee, C., Wu, S.S., and Chen, L.B.Photosensitization by 3,3'-dihexyloxacarbocyanine iodide: specific disruption of microtubules and inactivation of organelle motilityCancer Res.55(10)2063-2069(1995)
| Cas No. | 53213-82-4 | SDF | |
| 别名 | 碘代3,3'-二己氧基羰花青,DiOC6(3) iodide | ||
| Canonical SMILES | CCCCCCN1/C(OC2=CC=CC=C12)=C/C=C/C3=[N+](CCCCCC)C4=CC=CC=C4O3.[I-] | ||
| 分子式 | C29H37IN2O2 | 分子量 | 572.52 |
| 溶解度 | DMSO : ≥ 125 mg/mL (218.33 mM);Water : < 0.1 mg/mL (insoluble) | 储存条件 | Store at 2-8°C, protect from light |
| 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。 |
||
| 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 | 1.7467 mL | 8.7333 mL | 17.4666 mL |
| 5 mM | 0.3493 mL | 1.7467 mL | 3.4933 mL |
| 10 mM | 0.1747 mL | 0.8733 mL | 1.7467 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。
Nicotinamide improves in vitro lens regeneration in a mouse capsular bag model
Stem Cell Res Ther 2022 May 12;13(1):198.PMID:35550648DOI:10.1186/s13287-022-02862-8.
Background: Mammalian lens regeneration holds great potential as a cataract therapy. However, the mechanism of mammalian lens regeneration is unclear, and the methods for optimization remain in question. Methods: We developed an in vitro lens regeneration model using mouse capsular bag culture and improved the transparency of the regenerated lens using nicotinamide (NAM). We used D4476 and SSTC3 as a casein kinase 1A inhibitor and agonist, respectively. The expression of lens-specific markers was examined by real-time PCR, immunostaining, and western blotting. The structure of the in vitro regenerated lens was investigated using 3,3'-Dihexyloxacarbocyanine iodide \(DiOC6) and methylene blue staining, terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL), and transmission electron microscopy. Results: The in vitro lens regeneration model was developed to mimic the process of in vivo mammalian lens regeneration in a mouse capsular bag culture. In the early stage, the remanent lens epithelial cells proliferated across the posterior capsule and differentiated into lens fiber cells (LFCs). The regenerated lenses appeared opaque after 28 days; however, NAM treatment effectively maintained the transparency of the regenerated lens. We demonstrated that NAM maintained lens epithelial cell survival, promoted the differentiation and regular cellular arrangement of LFCs, and reduced lens-related cell apoptosis. Mechanistically, NAM enhanced the differentiation and transparency of regenerative lenses partly by inhibiting casein kinase 1A activity. Conclusion: This study provides a new in vitro model for regeneration study and demonstrates the potential of NAM in in vitro mammalian lens regeneration.
Targeting the Wnt/beta-catenin pathway in renal cell carcinoma
Anticancer Res 2014 Aug;34(8):4101-8.PMID:25075035doi
Background/aim: Advanced renal cancer still has a very poor prognosis. In this regard recent investigations demonstrated a constitutive activation of the Wnt signaling pathway in renal cell carcinoma (RCC) thereby promoting an exaggerated cell proliferation. Especially, β-catenin overactivation and the functional loss of endogenous Wnt antagonists are associated with RCC carcinogenesis and progression. Thus, influencing Wnt signaling might represent a promising target in RCC treatment. Materials and methods: It was recently confirmed that ethacrynic acid (EA), ciclopirox olamine (CIC) and piroctone olamine (PO) can inhibit Wnt signaling in various cancer cell lines. Herein we investigated their cytotoxic potential towards human RCC cells and their influence on the Wnt pathway concerning apoptosis as determined by 3,3'-Dihexyloxacarbocyanine iodide \(DiOC6) and propidium iodide (PI) staining in flow cytometry and immunoblotting. Results: All three agents, EA, CIC and PO triggered a significant apoptotic activity in tested RCC cell lines in a time- and concentration-dependent manner. Moreover, exposure to CIC and PO decreased the expression of β-catenin as the pivotal feature within the canonical Wnt pathway. However, β-catenin expression increased upon the treatment with EA. Conclusion: These results reveal a significant selective induction of apoptosis by EA, CIC and PO and suggest a suppression of RCC survival in part due to inhibition of Wnt/β-catenin signaling. The development of targeted-therapies affecting the Wnt signaling pathway might therefore lead to novel treatment options for RCC patients.
Granulocytes without degranulation: neutrophil function in granule-depleted cytoplasts
Proc Natl Acad Sci U S A 1983 Aug;80(16):4968-72.PMID:6308664DOI:10.1073/pnas.80.16.4968.
Neutrophils respond to a variety of stimuli by generating superoxide anion, degranulating, and aggregating. Because it has been suggested that fusion of granules with the plasmalemma (degranulation) is necessary for aggregation and superoxide anion generation, we have tested whether these responses can be demonstrated in "neutrophilic cytoplasts" (granule-free vesicles of cytoplasm enclosed by plasmalemma). When examined by electron microscopy, cytoplasts were found to be approximately 4 microns in diameter and essentially granule free. Cytoplasts exposed to fMet-Leu-Phe (0.1 microM) generated superoxide anion after a lag of 16 sec but released no detectable beta-glucuronidase, lysozyme, or elastase. Aggregation of cytoplasts, as measured by changes in light transmission, was also activated by fMet-Leu-Phe; no lag period was observed. Electron microscopy of the aggregates demonstrated clusters of cytoplasts with a scalloped appearance. Superoxide anion generation and aggregation of cytoplasts were also activated by phorbol 12-myristate 13-acetate, concanavalin A, and leukotriene B4. Exposure of cytoplasts to the dye 3,3'-Dihexyloxacarbocyanine iodide \(DiOC6(3)] led to dye uptake and enhancement of fluorescence, implying that the vesicles were sealed and maintained a membrane potential across the plasmalemma. Exposure of DiOC6(3)-loaded cytoplasts to fMet-Leu-Phe and PMA caused a rapid loss of dye fluorescence that was not inhibited by CN-, compatible with their lack of mitochondria. Exposure of dye-loaded cytoplasts to concanavalin A or leukotriene B4 caused an increase in fluorescence--i.e., a hyperpolarization. These results demonstrate that degranulation is not a prerequisite for aggregation or superoxide anion generation. The retention of ionic gradients and changes in membrane potential, as measured by DiOC6(3) fluorescence changes, suggest a fundamental role for ionic movements in activating superoxide anion generation and aggregation.
Mitochondrial alterations induced by 532 nm laser irradiation
Gen Physiol Biophys 2005 Jun;24(2):209-20.PMID:16118473doi
Mitochondrial alterations were monitored after low power green laser (532 nm, 30 mW) irradiation in the case of whole cells (B-14) and isolated mitochondria (from Wistar rat heart). 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium-bromide (MTT) assay products were significantly higher (by 8%) in irradiated B-14 cells as compared to non-irradiated controls. Mitochondrial transmembrane potential of B-14 cells, measured by means of a fluorescent probe 3,3'-Dihexyloxacarbocyanine iodide \(DiOC6(3)), significantly increased (by 13%) after exposure to green laser irradiation. Another MTT assay was used for isolated mitochondria suspensions in order to examine the effect of green laser irradiation on stimulation of processes related to oxidative phosphorylation. It revealed 31.3%-increase in MTT assay products in irradiated mitochondria as compared to controls. Laser irradiation of isolated mitochondria suspension did not significantly change 1,6-diphenyl-1,3,5-hexatriene (DPH) fluorescence anisotropy, indicating that mitochondrial membrane fluidity was not affected by laser light. Fluorescence emission spectra of irradiated as well as non-irradiated mitochondria suspensions showed fluorescence maximum at 635 nm, corresponding to emission of Protoporphyrin IX, which was significantly lower (by 20.7%) in irradiated sample.
Kinesin-mediated organelle translocation revealed by specific cellular manipulations
J Cell Biol 1994 Nov;127(4):1021-39.PMID:7962067DOI:10.1083/jcb.127.4.1021.
The distribution of membrane-bound organelles was studied in cultured hippocampal neurons after antisense oligonucleotide suppression of the kinesin-heavy chain (KHC). We observed reduced 3,3'-Dihexyloxacarbocyanine iodide \(DiOC6(3)) fluorescent staining in neurites and growth cones. In astrocytes, KHC suppression results in the disappearance of the DiOC6(3)-positive reticular network from the cell periphery, and a parallel accumulation of label within the cell center. On the other hand, mitochondria microtubules and microfilaments display a distribution that closely resembles that observed in control cells. KHC suppression of neurons and astrocytes completely inhibited the Brefeldin A-induced spreading and tubulation of the Golgi-associated structure enriched in mannose-6-phosphate receptors. In addition, KHC suppression prevents the low pH-induced anterograde redistribution of late endocytic structures. Taken collectively, these observations suggest that in living neurons, kinesin mediates the anterograde transport of tubulovesicular structures originated in the central vacuolar system (e.g., the endoplasmic reticulum) and that the regulation of kinesin-membrane interactions may be of key importance for determining the intracellular distribution of selected organelles.