5-Carboxyfluorescein diacetate N-succinimidyl ester
(Synonyms: 5-羧基二乙酸荧光素 N-琥珀酰亚胺酯) 目录号 : GC65544
5-Carboxyfluorescein diacetate N-succinimidyl ester 是一种细胞渗透性染料 (Ex=492 nm，Em=517 nm)。5-Carboxyfluorescein diacetate N-succinimidyl ester 可与细胞内分子共价结合，从而标记细胞。5-Carboxyfluorescein diacetate N-succinimidyl ester 可用于示踪淋巴细胞的迁移和增殖。
Cas No.:150206-05-6
Sample solution is provided at 25 µL, 10mM.
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Quality Control & SDS
- View current batch:
- Purity: >99.00%
- COA (Certificate Of Analysis)
- SDS (Safety Data Sheet)
- Datasheet
5-Carboxyfluorescein diacetate N-succinimidyl ester is a cell permeable dye (Ex=492 nm, Em=517 nm). 5-Carboxyfluorescein diacetate N-succinimidyl ester can label cells by covalently binding to intracellular molecules. 5-Carboxyfluorescein diacetate N-succinimidyl ester is used to track lymphocyte migration and proliferation[1].
Guidelines (Following is our recommended protocol. This protocol only provides a guideline, and should be modified according to your specific needs)[1].1. Dilute the stock solution in PBS (to give a 50 μM solution).2. Add 110 μL of this solution per milliliter of cells (to give a final concentration of 5 μM) and mix rapidly.3. After 5 min at room temperature, add 10 vol of PBS containing 5% FBS. 4. Centrifuge cells 5 min at 300 ×g, 20 ℃.5. Remove the supernatant, and wash threetimes, each time by resuspending in 10 vol PBS containing 5% FBS. 6. Centrifuge cells 5 min at 300 × g, 20 ℃, and removing the supernatant. Note: Labeling occurs rapidly, and it is essential that the tracker is dispersed as evenly and quickly as possible so that cells are uniformly labeled.
[1]. Christopher R Parish, et al. Use of the intracellular fluorescent dye CFSE to monitor lymphocyte migration and proliferation. Curr Protoc Immunol. 2009 Feb;Chapter 4:Unit4.9.
| Cas No. | 150206-05-6 | SDF | Download SDF |
| 别名 | 5-羧基二乙酸荧光素 N-琥珀酰亚胺酯 | ||
| 分子式 | C29H19NO11 | 分子量 | 557.46 |
| 溶解度 | DMSO : 20.83 mg/mL (37.37 mM; ultrasonic and warming and heat to 60°C) | 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 | 1.7939 mL | 8.9693 mL | 17.9385 mL |
| 5 mM | 0.3588 mL | 1.7939 mL | 3.5877 mL |
| 10 mM | 0.1794 mL | 0.8969 mL | 1.7939 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。
Immunosuppressive capabilities of mesenchymal stromal cells are maintained under hypoxic growth conditions and after gamma irradiation
Cytotherapy 2015 Feb;17(2):152-62.PMID:25453724DOI:10.1016/j.jcyt.2014.10.004
Background aims: The discovery of regenerative and immunosuppressive capacities of mesenchymal stromal cells (MSCs) raises hope for patients with tissue-damaging or severe, treatment-refractory autoimmune disorders. We previously presented a method to expand human MSCs in a bioreactor under standardized Good Manufacturing Practice conditions. Now we characterized the impact of critical treatment conditions on MSCs with respect to immunosuppressive capabilities and proliferation. Methods: MSC proliferation and survival after γ irradiation were determined by 5-Carboxyfluorescein diacetate N-succinimidyl ester and annexinV/4',6-diamidino-2-phenylindole (DAPI) staining, respectively. T-cell proliferation assays were used to assess the effect of γ irradiation, passaging, cryopreservation, post-thaw equilibration time and hypoxia on T-cell suppressive capacities of MSCs. Quantitative polymerase chain reaction and β-galactosidase staining served as tools to investigate differences between immunosuppressive and non-immunosuppressive MSCs. Results: γ irradiation of MSCs abrogated their proliferation while vitality and T-cell inhibitory capacity were preserved. Passaging and long cryopreservation time decreased the T-cell suppressive function of MSCs, and postthaw equilibration time of 5 days restored this capability. Hypoxic culture markedly increased MSC proliferation without affecting their T-cell-suppressive capacity and phenotype. Furthermore, T-cell suppressive MSCs showed higher CXCL12 expression and less β-galactosidase staining than non-suppressive MSCs. Discussion: We demonstrate that γ irradiation is an effective strategy to abrogate MSC proliferation without impairing the cells' immunosuppressive function. Hypoxia significantly enhanced MSC expansion, allowing for transplantation of MSCs with low passage number. In summary, our optimized MSC expansion protocol successfully addressed the issues of safety and preservation of immunosuppressive MSC function after ex vivo expansion for therapeutic purposes.