Acridine Orange
(Synonyms: 吖啶橙,NSC 194350) 目录号 : GC41242
Acridine Orange是一种可穿透细胞的核酸选择性荧光染料。
Cas No.:494-38-2
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: >75.00%
- 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Acridine Orange染色液
（1）Acridine Orange染料储存液: 使用DMSO将Acridine Orange溶解成1mg/ml的储存液。
注意:未使用的储存液建议分装后于-4℃或-20℃避光保存（在氮气下储存），避免反复冻融。
（2）Acridine Orange染料工作液制备: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0.5-20μg/ml的Acridine Orange工作液。
注意: 请根据实际情况调整工作液浓度，现用现配
2、细胞的DNA和RNA染色：
(1) 收集细胞悬液，将约0.2ml/份的原始细胞悬液转移至2/5 mL体积的小玻璃或塑料管中, 放置在冰上冷却；
(2) 缓慢加入0.4 mL预冷的细胞透化液，将细胞置于冰上静置约15s；
(3) 缓慢加入1.2 mL预冷的Acridine Orange工作液，冰上避光孵育2-15min；
(4) 使用流式细胞仪测量并记录细胞荧光，在488 nm处激发，通过发射滤光片和分色镜区分绿色荧光（在515-545 nm处测量）处的dsDNA和红色发光（优选在640或650nm以上测量）处的RNA，Acridine Orange结合ssDNA后的激发光/发射光波长为460/650nm。
3、细胞贴壁染色（冰上操作）
（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染料工作液，轻轻晃动使染料均匀覆盖所有细胞，冰上避光孵育2-15min；
（4）吸弃染料工作液，使用培养液清洗盖玻片2~3次，通过荧光显微镜进行观察。
4、显微镜检测：使用共聚焦激光扫描显微镜（如FV3000，奥林巴斯）观察细胞，或者通过MetaMorph对每个帧中Acridine Orange的总荧光进行定量。当与dsDNA结合、与ssDNA或RNA结合以及酸性条件下时，Acridine Orange分别表现出502/525、460/650和475/590 nm的激发光/发射光谱。
注意事项：
①细胞如需固定后染色，建议使用70%乙醇进行固定，冰上固定≥2小时，固定后使用预冷的PBS漂洗1-2次，除去所有乙醇，而后按照上述步骤进行染色；
②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；
为了您的安全和健康，请穿实验服并戴一次性手套操作。
Acridine orange is a cell-permeable, nucleic acid-selective fluorescent cationic dye useful for cell cycle determination and detection of cellular autophagy. [1][2][3] It exhibits excitation/emission spectra of 502/525, 460/650, and 475/590 nm when bound to dsDNA, bound to ssDNA or RNA, and under acidic conditions, respectively. [1] [2] The ratio of acridine orange emission at 590 to emission at 525 nm can be used to quantify the increase in the number of acidic vesicular organelles observed during cellular autophagy.
Reference:
[1]. Virant-Klun, I., Tomazevic, T., and Meden-Vrtovec, H. Sperm single-stranded DNA, detected by acridine orange staining, reduces fertilization and quality of ICSI-derived embryos. J.Assist.Reprod.Genet. 19(7), 319-328 (2002).
[2]. Han, J., and Burgess, K. Fluorescent indicators for intracellular pH. Chem. Revs. 110(5), 2709-2728 (2010).
[3]. Thomé, M.P., Filippi-Chiela, E.C., Villodre, E.S., et al. Ratiometric analysis of Acridine Orange staining in the study of acidic organelles and autophagy. J. Cell Sci. 129(24), 4622-4632 (2016).
| Cas No. | 494-38-2 | SDF | |
| 别名 | 吖啶橙,NSC 194350 | ||
| 化学名 | N3,N3,N6,N6-tetramethyl-3,6-acridinediamine | ||
| Canonical SMILES | CN(C)C1=CC(N=C(C=C(N(C)C)C=C2)C2=C3)=C3C=C1 | ||
| 分子式 | C17H19N3 | 分子量 | 265.4 |
| 溶解度 | 0.3mg/mL in ethanol, 20mg/mL in DMSO, 2mg/mL in DMF | 储存条件 | Store at -20°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 | 3.7679 mL | 18.8395 mL | 37.679 mL |
| 5 mM | 0.7536 mL | 3.7679 mL | 7.5358 mL |
| 10 mM | 0.3768 mL | 1.8839 mL | 3.7679 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。
Nonyl Acridine Orange as a Prospective Photocatalyst in Chalcogenylation of Coumarins and Quinolinones
J Org Chem 2022 Nov 18;87(22):15261-15272.PMID:36310352DOI:10.1021/acs.joc.2c01803.
A mild and efficient method for preparation of 3-sulfenyl and 3-selenyl coumarins and quinolinones mediated by artificial light or sunlight is presented. The elaborated protocol highlights the use of nonyl Acridine Orange as a photocatalyst to generate a sulfenyl radical from thiols that is further trapped by a heterocycle. The utility of the protocol is justified by a diverse scope of thiols, including short cysteine-containing peptides. The same reaction conditions can be applied for preparation of 3-selenyl coumarins and quinolinones. Various protected and unprotected selenocysteine-containing peptides were successfully utilized demonstrating high tolerance for amino acids with sensitive groups (Arg, Lys, Trp, His, and Tyr).
Comparison of Acridine Orange and giemsa stains for malaria diagnosis
Korean J Parasitol 1995 Dec;33(4):391-4.PMID:8591019DOI:10.3347/kjp.1995.33.4.391.
Recently, the Acridine Orange (AO) staining method has improved for identification of malaria parasites. Fixed and preserved blood smears of malaria patients were used for comparative analysis of AO and Giemsa stains. The AO staining method required less time and was more sensitive under lower magnification than the Giemsa staining method. The AO staining method provides an alternative to Giemsa for malaria diagnosis in the field and laboratory.
Acridine Orange interaction with DNA: Effect of ionic strength
Biochim Biophys Acta Gen Subj 2017 Apr;1861(4):900-909.PMID:28130157DOI:10.1016/j.bbagen.2017.01.023.
Background: The study of Acridine Orange (AO) spectral characteristics and the quenching of its singlet and triplet excited states by TEMPO radical at its binding to DNA in the function of the DNA concentration and in the absence and presence of NaCl is reported. Methods: The study was performed using steady-state and time resolved optical absorption and florescence, fluorescence correlation spectroscopy and resonant light scattering techniques. Results: The presence of different species in equilibrium: AO monomers and aggregates bound to DNA, has been demonstrated, their relative content depending on the DNA and the AO concentrations. At high DNA concentration the AO monomers are protected against the contact with other molecules, thus reducing the AO excited state quenching. The addition of NaCl reduces the AO binding constant to DNA, thus reducing the AO and DNA aggregation. Conclusions: The interaction of AO with DNA is a complex process, including aggregation and disaggregation of both components. This modifies the AO excited state characteristics and AO accessibility to other molecules. The salt reduces the DNA effects on the AO excited state characteristics thus attenuating its effects on the AO efficacy in applications. General significance: This study demonstrates that the interaction of photosensitizers with DNA, depending on their relative concentrations, can both decrease and increase the photosensitizer efficacy in applications. The salt is able to attenuate these effects.
Acridine Orange as a coreactant for efficient electrogenerated chemiluminescence of tris(2,2'-bipyridine)ruthenium(ii) and its use in selective and sensitive detection of thiourea
Chem Commun (Camb) 2020 May 11;56(38):5154-5157.PMID:32255109DOI:10.1039/d0cc01273d.
A new and efficient anodic Ru(bpy)32+ ECL system by using Acridine Orange (3,6-Bis(dimethylamino)acridine, AO) as a coreactant was reported in a neutral medium. The developed Ru(bpy)32+-AO system displayed a higher ECL intensity than that of the classic Ru(bpy)32+- oxalate ECL system, and was further exploited for the ECL detection of thiourea for the first time.
Impact of Acridine Orange in Patients With Local Recurrent Soft Tissue Sarcoma
In Vivo 2020 Sep-Oct;34(5):2745-2750.PMID:32871809DOI:10.21873/invivo.12097.
Background/aim: Local recurrence in soft tissue sarcoma (STS) is a risk factor of worse prognosis. Although a few studies have shown that adjuvant therapy with Acridine Orange (AO) is effective for local control of primary STS, there have been no reports examining its effectiveness for local recurrence. Patients and methods: This retrospective study included 36 patients with first local recurrence of STS. Of them, 23 patients received wide excision without AO therapy (Wide group); the other 13 patients received marginal excision with AO therapy (AO group). We compared re-recurrence rates between these two groups. Results: The total re-recurrence rate was 43.5% in the Wide group and 46.2% in the AO group. There was no significant difference in local re-recurrence-free survival and overall survival between the two groups. Conclusion: Adjuvant AO therapy combined with a marginal excision suppresses local re-recurrence rates of individuals with local STS recurrence.