TO-PRO 1
目录号 : GC64792
TO-PRO 1是一种具有单一阳离子侧链和低碱选择性的绿色荧光染料，与核酸结合后的最大激发/发射光为515/531 nm。
Cas No.:157199-59-2
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
本方案仅提供一个指导，请根据您的具体需要进行修改。
1、配制工作液: 使用合适的缓冲液（如HHBS或PBS）稀释浓度为5 mM的TO-PRO 1贮存液，配置成1-5μM的工作液。
注意:
①首次使用储存液时建议根据单次用量分装成小规格，-20℃或-80℃避光冻存，避免反复冻融；
②建议初次实验时设置染色液浓度梯度，以确定最佳工作浓度；
③高染料浓度可能引起其它细胞结构的非特异染色；
④请根据实际情况调整工作液浓度，现用现配。
2、细胞染色（以6孔板为例）
2.1细胞悬浮染色
（1）悬浮细胞经1000g离心3-5min。弃去上清液，使用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经37 °C预热的TO-PRO 1染料工作液重悬细胞，37 °C避光孵育15-60min分钟，不同细胞最佳培养时间不同。
（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。
（5）使用缓冲液重悬细胞，通过荧光显微镜或流式细胞技术进行观察。
2.2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，37 °C避光孵育15-60min分钟。
（4）吸弃染料工作液，使用培养液洗盖玻片2~3次，通过荧光显微镜进行观察。
3.显微镜检测：TO-PRO 1结合核酸后的最大激发/发射光为515/531 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。
References:
[1]. Elena Gorokhova, Lisa Mattsson, Annica M Sundström. A comparison of TO-PRO-1 iodide and 5-CFDA-AM staining methods for assessing viability of planktonic algae with epifluorescence microscopy. 2012 Jun;89(3):216-21. doi: 10.1016/j.mimet.2012.03.005. Epub 2012 Mar 14.
TO-PRO 1 is a monomeric cyanine dye with a single cationic side chain and low base selectivity that is able to enter cells with damaged membranes and bind to nucleic acids, producing bright green fluorescence. In addition, TO-PRO 1 shows excellent staining results for diatoms that are challenging to stain. TO-PRO 1 is almost non-fluorescent when not bound to nucleic acids, but shows strong fluorescence when bound to nucleic acids, with a maximum excitation/emission light of 515/531 nm[1].
TO-PRO 1是一种具有单一阳离子侧链和低碱选择性的单体花菁染料，能够进入膜受损的细胞并与核酸结合，产生明亮的绿色荧光。此外，TO-PRO 1对染色具有挑战性的硅藻表现非常出色的染色效果。TO-PRO 1未结合核酸时几乎不发荧光，但当与核酸结合后表现出强烈荧光，最大激发/发射光为515/531 nm[1]。
References:
[1]. Elena Gorokhova , Lisa Mattsson, Annica M Sundström. A comparison of TO-PRO-1 iodide and 5-CFDA-AM staining methods for assessing viability of planktonic algae with epifluorescence microscopy. 2012 Jun;89(3):216-21. doi: 10.1016/j.mimet.2012.03.005. Epub 2012 Mar 14.
| Cas No. | 157199-59-2 | SDF | Download SDF |
| 分子式 | C24H29I2N3S | 分子量 | 645.38 |
| 溶解度 | 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 | 1.5495 mL | 7.7474 mL | 15.4947 mL |
| 5 mM | 0.3099 mL | 1.5495 mL | 3.0989 mL |
| 10 mM | 0.1549 mL | 0.7747 mL | 1.5495 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。
Fluorochrome-functionalized nanoparticles for imaging DNA in biological systems
ACS Nano 2013 Mar 26;7(3):2032-41.PMID:23373524DOI:10.1021/nn305962n.
Attaching DNA binding fluorochromes to nanoparticles (NPs) provides a way of obtaining NPs that bind to DNA through fluorochrome mediated interactions. To obtain a nanoparticle (NP) that bound to the DNA in biological systems, we attached the DNA binding fluorochrome, TO-PRO 1 (TO), to the surface of the Feraheme (FH) NP, to obtain a fluorochrome-functionalized NP denoted TO-FH. When reacted with DNA in vitro, TO-FH formed microaggregates that were characterized by fluorescence, light scattering, and T2 changes. The formation of DNA/TO-FH microaggregates was also characterized by AFM, with microaggregates exhibiting a median size of 200 nm, and consisting of DNA and multiple TO-FH NPs whose individual diameters were only 25-35 nm. TO-FH failed to bind normal cells in culture, but treatment with chemotherapeutic agents or detergents yielded necrotic cells that bound TO-FH and vital fluorochromes similarly. The uptake of TO-FH by HT-29 xenografts (treated with 5-FU and oxaliplatin) was evident by surface fluorescence and MRI. Attaching multiple DNA binding fluorochromes to magnetic nanoparticles provides a way of generating DNA binding NPs that can be used to detect DNA detection by microaggregate formation in vitro, for imaging the DNA of necrotic cells in culture, and for imaging the DNA of a tumor treated with a chemotherapeutic agent. Fluorochrome functionalized NPs are a multimodal (magnetic and fluorescent), highly multivalent (n ≈ 10 fluorochromes/NP) nanomaterials useful for imaging the DNA of biological systems.
Fluorescent properties of oligonucleotide-conjugated thiazole orange probes
Photochem Photobiol 2002 Mar;75(3):201-10.PMID:11950085DOI:10.1562/0031-8655(2002)075<0201:fpooct>2.0.co;2.
The fluorescence properties of thiazole orange, linked via a (1) hydrophobic alkyl or a (2) hydrophilic ethylene glycol chain to the central internucleotidic phosphate group of a pentadeca-2'-deoxyriboadenylate (dA15), are evaluated. Linkage at the phosphate group yields two stereoisomers, S-isomer of the phosphorus chiral center (Sp) and R-isomer of the phosphorus chiral center (Rp); these are studied separately. The character of the linkage chain and the chirality of the internucleotidic phosphate linkage site influence the fluorescent properties of these thiazole orange-oligonucleotide conjugates (TO-probes). Quantum yields of fluorescence (phifl) of between 0.04 and 0.07 were determined for the single-stranded conjugates. The fluorescence yield increased by up to five times upon hybridization with the complementary sequence (d5'[CACT15CAC3']); (phifl values of between 0.06-0.35 were determined for the double-stranded conjugates. The phifl value (0.17) of thiazole orange, 1-(N,N'-trimethylaminopropyl)-4-[3-methyl-2,3-dihydro-(benzo-1,3-thiazole)-2-methylidene]-quinolinium iodide (TO-PRO 1) in the presence of the oligonucleotide duplex (TO-PRO 1: dA15.d5'[CACT15CAC3'] (1:1)) is much less than that for some of the hybrids of the conjugates. Our studies, using steady-state and time-resolved fluorescence experiments, show that a number of discrete fluorescent association species between the thiazole orange and the helix are formed. Time-resolved studies on the four double-stranded TO-probes revealed that the fluorescent oligonucleotide-thiazole orange complexes are common, only the distribution of the species varies with the character of the chain and the chirality at the internucleotidic phosphate site. Those TO-probes in which the isomeric structure of the phosphate-chain linkage is Rp, and therefore such that the fluorophore is directed toward the minor groove, have higher phifl values than the Sp isomer. Of the systems studied, thiazole orange linked by an alkyl chain to the internucleotidic phosphate (Rp isomer) has the highest phifl and the greatest fraction of the longest-lived fluorescent thiazole orange species (in the hybrid form).