4-Di-10-ASP
目录号 : GC680014-Di-10-ASP 是一种荧光亲脂性示踪剂 (Excitation 485 nm; Emission 620 nm)。4-Di-10-ASP 可以用来对磷脂膜进行特殊染色。
Cas No.:95378-73-7
Sample solution is provided at 25 µL, 10mM.
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Quality Control & SDS
- View current batch:
- Purity: >98.00%
- COA (Certificate Of Analysis)
- SDS (Safety Data Sheet)
- Datasheet
4-Di-10-ASP is a fluorescent lipophilic tracer (Excitation 485 nm; Emission 620 nm). 4-Di-10-ASP can be used to stain phospholipid membranes in a specific manner[1][2].
Guidelines (Following is our recommended protocol. This protocol only provides a guideline, and should be modified according to your specific needs).
1. 4-Di-10-ASP (1 μM), DOPC (10 mM), and DOPG (1 mM) is dissolved in methanol/chloroform (25 L, 1:2 v/v).
2. The solution is allowed to dry overnight under vacuum to obtain lamellar lipid films, which in turn are hydrated with the transcription/translation solution (25 L) for three hours at 37℃.
3. An aliquot (10 L) of the solution thus prepared is placed on a glass slide and sealed by a cover glass.
4. The sample is immediately observed with a confocal laser-scanning microscope, an argon laser (488 nm) is employed to excite the 4-Di-10-ASP[2].
[1]. Z J Huang, et al. Partition coefficients of fluorescent probes with phospholipid membranes. Biochem Biophys Res Commun. 1991 Nov 27;181(1):166-71.
[2]. Shin-ichiro M Nomura, et al. Gene expression within cell-sized lipid vesicles. Chembiochem. 2003 Nov 7;4(11):1172-5.
| Cas No. | 95378-73-7 | SDF | Download SDF |
| 分子式 | C34H55IN2 | 分子量 | 618.72 |
| 溶解度 | DMSO : 33.33 mg/mL (53.87 mM; ultrasonic and warming and heat to 60°C) | 储存条件 | 4°C, away from moisture and 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。 |
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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 | 1.6162 mL | 8.0812 mL | 16.1624 mL |
| 5 mM | 0.3232 mL | 1.6162 mL | 3.2325 mL |
| 10 mM | 0.1616 mL | 0.8081 mL | 1.6162 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。
Partition coefficients of fluorescent probes with phospholipid membranes
Biochem Biophys Res Commun 1991 Nov 27;181(1):166-71.PMID:1958185DOI:10.1016/s0006-291x(05)81396-8.
A method for determination of membrane partition coefficients of five fluorescent membrane probes, 1,6-diphenyl-1,3,5-hexatriene (DPH), p-((6-phenyl)-1,3,5-hexatrienyl) benzoic acid (DPH carboxylic acid), 3-(p-(6-phenyl)-1,3,5-hexatrienyl)phenylpropionic acid (DPH propionic acid), 1-(4-trimethylammoniumphenyl)-6-phenyl-1,3,5-hexatriene (TMA-DPH) and N-4-(4-didecylaminostyryl)-N-methylpyridinium iodide (4-Di-10-ASP), was developed utilizing the fluorescence enhancement of a constant probe concentration by titration with excess phospholipid liposomes. The partition coefficients of DPH, DPH carboxylic acid, DPH propionic acid, TMA-DPH and 4-Di-10-ASP into dipalmitoylphosphatidylcholine membranes were determined to be 1.3.10(6), 1.0.10(6), 6.5.10(5), 2.4.10(5) and 2.8.10(6) respectively. Knowledge of the partition coefficients may help select a lipid concentration for membrane studies that necessitate a probe's dominant incorporation into membranes.
Effects of increased intraocular pressure on rat retinal ganglion cells
Int J Dev Neurosci 2001 Apr;19(2):209-18.PMID:11255034DOI:10.1016/s0736-5748(00)00073-3.
The effects of elevated intraocular pressure (IOP) on the morphology of rat retinal ganglion cells (RGCs) was analyzed in this study. After cauterizing two limbal derived episcleral veins, IOP in experimental eyes was elevated 1.5--1.8 times that of control. RGCs of experimental and control eyes were analyzed after: bilateral tectal injections of Fluoro-Gold, and application of fluorescent dye crystals, 4-Di-10-ASP to the proximal stump of the cut optic nerve, at different time intervals after IOP elevation. The RGCs in control and experimental eyes were evaluated at 4, 6, 8, and 10 weeks by counting, as well as by determining the soma diameter. The dendritic field of three types (I, II, III) of RGCs between control and experimental eyes were also studied at 4,6,10 weeks after IOP elevation. At every time point, the number of cells in experimental eyes were significantly less than those of the control eyes. The average retinal ganglion cell death was 3--4% per week in the eyes with elevated IOP. The soma and dendritic field diameter of the RGCs in the experimental eyes were significantly larger in all cell types. However, types I and III cells expanded their dendritic fields more rapidly than type II cells. Furthermore, dendritic fields of surviving RGCs in experimental eyes occupied about the same extent of the retina as the controls. The increase in soma diameter and expansion of dendritic fields in the remaining RGCs in eyes with elevated IOP suggests the existence of plasticity in adult retina.
Sema-3A indirectly disrupts the regeneration process of goldfish optic nerve after controlled injury
Graefes Arch Clin Exp Ophthalmol 2010 Oct;248(10):1423-35.PMID:20449604DOI:10.1007/s00417-010-1377-y.
Background: Neurons of adult mammalian CNS are prevented from regenerating injured axons due to formation of a non-permissive environment. The retinal ganglion cells (RGC), which are part of the CNS, share this characteristic. In sharp contrast, the RGC of lower vertebrates, such as fish, are capable of re-growing injured optic nerve axons, and achieve, through a complex multi-factorial process, functional vision after injury. Semaphorin-3A (sema-3A), a member of the class 3 semaphorins known for its repellent and apoptotic activities, has previously been shown to play a key role in the formation of a non-permissive environment after CNS injury in mammalians. Methods: The expression of sema-3A and its effect on regenerative processes in injured gold fish retina and optic nerve were investigated in this study. Unilateral optic nerve axotomy or crush was induced in goldfish. 2 microl sema-3A was injected intraviterally 48 hours post injury. Neuronal viability was measured using the lipophilic neurotracer dye 4-Di-10-ASP. Axonal regeneration was initiated using the anterograde dye dextran. Retinas and optic nerves were collected at intervals of 2, 3, 7, 14 and 28 days after the procedure. Using Western blot and immunohistochemical analysis, the expression levels of semaphorin-3A, axonal regeneration, the removal of myelin debris and macrophage invasion were studied. Results: We found a decrease in sema-3A levels in the retina at an early stage after optic nerve injury, but no change in sema-3A levels in the injured optic nerve. Intravitreal injection of sema-3A to goldfish eye, shortly after optic nerve injury, led to destructive effects on several pathways of the regenerative processes, including the survival of retinal ganglion cells, axonal growth, and clearance of myelin debris from the lesion site by macrophages. Conclusions: Exogenous administration of sema-3A in fish indirectly interferes with the regeneration process of the optic nerve. The findings corroborate our previous findings in mammals, and further validate sema-3A as a key factor in the generation of a non-permissive environment after transection of the optic nerve.
Expression of N-methyl-d-aspartate receptor 1 in rats with chronic ocular hypertension
Neuroscience 2007 Nov 23;149(4):908-16.PMID:17942238DOI:10.1016/j.neuroscience.2007.07.056.
High levels of glutamate can be toxic to retinal ganglion cells (RGCs). This study investigated the relationship between the N-methyl-d-aspartate receptor 1 (NR) and RGC death in a rat model of chronic ocular hypertension (COHT). COHT was induced in one eye of each rat by episcleral vein cauterization. Retinal protein expression was evaluated at 1, 3, 5 and 9 weeks after cauterization. Quantitative real-time polymerase chain reaction and Western blot analysis showed that NR1 expression was significantly increased in cauterized retinae. NR1 immunoreactivity was observed in the inner nuclear layer (INL) and ganglion cell layer (GCL) in the retina of rats with COHT. RGC density was evaluated after retrograde labeling with fluoro-gold (FG) and 4-Di-10-ASP (DiA). A significant decrease in RGC density was observed in ocular hypertensive eyes, and NR1 expression in the GCL suggested an important role of NR1 in the death of RGCs. Memantine (10 mg/kg), an N-methyl-d-aspartate receptor antagonist, was administered orally once daily for up to 5 weeks, while rats in the control group received vehicle phosphate-buffered saline only. Treatment with memantine resulted in a significant reduction in RGC loss and NR1 expression in the eyes of rats COHT. These findings suggest that excessive expression of NR1 is involved in RGC death in glaucoma.
Up-regulation of semaphorin expression in retina of glaucomatous rabbits
Graefes Arch Clin Exp Ophthalmol 2003 Aug;241(8):673-81.PMID:12827374DOI:10.1007/s00417-003-0684-y.
Background: Glaucoma is a term encompassing a variety of diseases that end in the death of retinal ganglion cells (RGC). Although a variety of factors can initiate the disease onset, increased intraocular pressure (IOP) is one of the major risk factors. In our previous study we found that semaphorins were causally involved in RGC death following axotomy. Since a common feature of all retinal neuropathies is axonal damage, we hypothesized that semaphorins are involved in glaucoma-induced RGC death. The purpose of this study was to analyze the effect of increased IOP on RGC viability and to analyze semaphorin expression pattern in glaucomatous retinas. Methods: Utilizing retrograde-labeled dye (4-Di-10-ASP) and hematoxylin-eosin staining, we investigated the effect of elevated levels of IOP on RGC viability. In addition, immunohistochemical analysis and western blotting were used to study the pattern of semaphorin expression in retinas of rabbits with genetically developed increased IOP and subsequently glaucoma. Results: Using specific anti-semaphorin antibodies, the expression of a single protein with the size of a semaphorin protein, 110 kDa, was detected; its expression was up-regulated in glaucomatous rabbits compared with controls. Time-course analysis revealed that semaphorin expression peaked between 2 and 6 months of age and declined thereafter. Immunohistochemical analysis revealed that semaphorin expression was up-regulated specifically in the ganglion cell layer, which is a structure that is highly affected in glaucoma. Conclusion: Deciphering the molecular mechanisms of glaucoma-induced death and its mediators is a crucial step towards designing new therapeutic strategies to treat this incurable disease.