| Overview |
 Printer Friendly Version
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| Ex/Em (nm) | 445/503 |
| MW | ~350 |
| CAS # | N/A |
| Solvent | N/A |
| Storage | F/D/L |
| Category |
Cell Biology Labeling Cells |
| Related |
Fluorescence Imaging Biochemical Assays |
| Spectrum | Advanced Spectrum Viewer |
1. Prepare Lysosome-staining solution:
1.1 Warm LysoBrite™ dyes to room temperature.
1.2 Prepare dye working solution by diluting 20 µL of 500 X LysoBrite™ dyes to 10 mL of Hanks and 20 mm Hepes buffer or buffer (HBSS) of your choice.
Note 1: 20 µL of LysoBrite™ dye is enough for one 96-well plate. Aliquot and store unused LysoBrite™ dye stock solutions at < -15 oC. Protect it from light and avoid repeated freeze-thaw cycles.
Note 2: The optimal concentration of the fluorescent lysosome indicator varies depending on the specific application. The staining conditions may be modified according to the particular cell type and the permeability of the cells or tissues to the probe.
2. Prepare and stain cells:
2.1 For adherent cells: a). Grow cells either in a 96-well black wall/clear bottom plate (100 µL/well/96-well plate) or on cover-slips inside a petri dish filled with the appropriate culture medium. When cells reach the desired confluence, add equal volume of the dye-working solution (from Step 1.2). b). Incubate the cells in a 37 °C, 5% CO2 incubator for 30 minutes. c). Wash the cells twice with pre-warmed (37 °C) Hanks and 20 mM Hepes buffer (HBSS) or buffer of your choice, fill the cell wells with HBSS or growth medium. d). Observe the cells using a fluorescence microscope fitted with a desired filter set.
Note: It is recommended to increase either the labeling concentration or the incubation time to allow the dye to accumulate if the cells do not appear to be sufficiently stained.
2.2 For suspension cells: a). Add equal volume of dye-working solution (from Step 1.2) into the cells. Incubate the cells in a 37 °C, 5% CO2 incubator for 30 minutes. b). Wash the cells twice with pre-warmed (37 °C) Hanks and 20 mM Hepes buffer (HBSS) or buffer of your choice, fill the cell wells with HBSS or growth medium. c). Observe the cells using a fluorescence microscope equipped with a desired filter set.
Note 1: It is recommended to increase either the labeling concentration or the incubation time to allow the dye to accumulate if the cells do not appear to be sufficiently stained.
Note 2: Suspension cells may be attached to cover-slips that have been treated with BD Cell-Tak® (BD Biosciences) and stained as adherent cells (see Step 2.1).
| References & Citations |
 Citation Explorer
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Autophagy proteins are not universally required for phagosome maturation
Authors: Marija Cemma, Sergio Grinstein, John H Brumell
Journal: Autophagy (2016): 1440--1446
Differential detection of tumor cells using a combination of cell rolling, multivalent binding, and multiple antibodies
Authors: Ja Hye Myung, Khyati A Gajjar, Jihua Chen, Robert E Molokie, Seungpyo Hong
Journal: Analytical chemistry (2014): 6088--6094
Versatile fabrication of nanoscale sol--gel bioactive glass particles for efficient bone tissue regeneration
Authors: Bo Lei, Xiaofeng Chen, Xue Han, Jiaan Zhou
Journal: Journal of Materials Chemistry (2012): 16906--16913
Advanced glycation end-products increase IL-6 and ICAM-1 expression via RAGE, MAPK and NF-$kappa$B pathways in human gingival fibroblasts
Authors: K Nonaka, Y Kajiura, M Bando, E Sakamoto, Y Inagaki, JH Lew, K Naruishi, T Ikuta, K Yoshida, T Kobayashi
Journal: Journal of Periodontal Research
AAT Bioquest AAT Bioquest是一家位于美国的生物公司，前身为ABD Bioquest，总部位于加利福尼亚州。专门从事光学检测技术十多年，一直致力于光谱学检测领域技术的创新和突破。其独特的光学检测技术，综合了化学、生物学和信息学等各个领域的研究，引领了比色、荧光和发光技术新一代光学探针的浪潮。AAT Bioquest在全球拥有强大的经验丰富的专业分销商网络，为从小型研究机构到《财富》500强企业的各类客户提供卓越的产品和定制服务。
美国AATBioquestInc.(前身是ABDBioquest，Inc.)是一家为从事生命科学研究、诊断研发及药物开发的科学家研发、生产和销售生物分析研究试剂和试剂盒的公司。公司致力于光谱学检测领域，包括吸收（颜色），荧光和发光技术。AATBioquest的产品帮助全世界的科学家和生物医药研究者更好的了解生物化学，免疫学，细胞生物学和分子生物学等领域。AATBioquest会不断介绍新产品，快速的丰富各个领域的产品。
1）我们提供反应荧光探针和发光探针，生物素和端粒酶能够应用于标记药物小分子和生物聚合物，如蛋白、核酸以及其他碳水化合物；2）我们研究并生产荧光和发光探针用于检测蛋白，核酸和活细胞。3）我们不断的推出新型的荧光和发光探针用于检测多种酶，特别是检测水解酶和氧化还原酶类；4）我们致力于开发用于信号转导研究的试剂；5）我们提供生理和神经探针，特别是钙离子指示剂和膜电位探针。
作为AATBioquestInc.的中国区域代理，艾美捷科技为中国客户提供光谱学检测领域，包括吸收（颜色），荧光和发光技术等全系列解决方案。我们也将一如既往更加努力为国内用户提供快捷、方便的高质量产品，同时更为您售前售后全面技术支持。
AATBioquest,Inc.(formerlyABDBioquest,Inc.)develops,manufacturesandmarketsbioanalyticalresearchreagentsandkitstoscientistsengagedinlifesciencesresearch,diagnosticR&Danddrugdiscovery.Wespecializeintheareaofphotometricdetectionsincludingabsorption(color),fluorescenceandluminescencetechnologies.TheCompany"sproductsenablescientistsandbiomedicalresearcherstobetterunderstandbiochemistry,immunology,cellBIOLOGyandmolecularbiology.AATBioquestconstantlyintroducesnewproducts,andoffersarapidlyexpandinglistofproductsthataregroupedintoseveralproductlines.
1)Ourreactivefluorescentandluminescentprobes,biotinsandtagenzymesareusedforlabelingsmalldrugmoleculesandbiopolymers,e.g,proteins,nucleicacidsandcarbohydrates;2)Wedevelopfluorescentandluminescentprobesfordetectingproteins,nucleicacidsandlivecells;3)Weconstantlyintroducenovelfluorescentandluminescentprobesfordetectingvariousenzymes,inparticular,hydrolyticandredoxenzymes;4)Wefocusondevelopingreagentsforsignaltransductionresearch;and5)Wealsoofferphysiologicalandneurologicalprobes,e.g.,calciumindicatorsandmembranepotentialprobes.
Besidesthestandardcatalogproductswealsooffercustomservicetomeetyourspecialresearchneeds.Ourcurrentservicesincludecustomsynthesisofcolorimetric,fluorescentandluminescentprobes,customdevelopmentofbiochemical,cell-basedanddiagnosticassaysandcustomscreeningofyourcompoundlibrariesagainstyourdefinedtargetsusingourvalidatedHTSassays.
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分子生物学实验的染料主要涉及到核酸染料和蛋白质染料.核酸染料主要有EB（溴化乙锭,高致癌性）,goldview,sybr green（实时定量PCR时常用染料）.这些染料可以和核酸双链分子特异性结合发出强荧光而被检测到.蛋白质染料最常用的是考马斯亮蓝 R-250,硝酸银.其中硝酸银有时也用于核酸染色.
染料分为天然染料和人工染料两种。天然染料有胭脂虫红、地衣素、石蕊和苏木素等，它们多从植物体中提取得到，其成分复杂，有些至今还未搞清楚。目前主要采用人工染料，也称煤焦油染料，多从煤焦油中提取获得，是苯的衍生物。多数染料为带色的有机酸或碱类，难溶于水，而易溶于有机溶剂中。为使它们易溶于水，通常制成盐类。
染料可按其电离后染料离子所带电荷的性质，分为酸性染料、碱性染料、中性（复合）染料和单纯染料四大类。 标本干燥后即进行固定，固定的目的有三个：
1）杀死微生物，固定细胞结构。
2）保证菌体能更牢的粘附在载玻片上，防止标本被水冲洗掉。
3）改变染料对细胞的通透性，因为死的原生质比活的原生质易于染色。
产品主要应用：点击化学(Clickchemistry)、蛋白质组学研究中的双向荧光差异凝胶电泳(2DDIGE)和实时荧光定量PCR(RealtimePCR)。
菁染料是性能优良的荧光标记染料，摩尔吸光系数在荧光染料中是最高的。N-羟基琥珀酰亚胺酯是最常用的脂肪氨基标记试剂，广泛用于蛋白质、氨基酸多肽、抗体、核酸及其他生物分子的标记和检测。通过改变次甲基链的长度,可改变其荧光发射波长，每增加一个双键，按照Huoffman规则正好红移约100nm。
菁染料Cy3和Cy5已成为基因芯片的首选荧光标记物；另外,Cy5,Cy5.5和Cy7，Cy7.5的吸收在近红外区背景非常低，是荧光强度最高、最稳定的长波长染料，特别适合于活体小动物体内成像。但由于菁染料,尤其是不对称菁染料的合成副反应多,副产物极性相近,产物的分离提纯相当困难。菁染料特别是水溶性菁染料分子极性大,分离提纯越加困难。Lumiprobe供应脂溶性和水溶性菁染料。
相关产品：
产品分子结构可替代染料编号：AGF1371A
6-ROX-N-羟基琥珀酰亚胺酯
ROXNHSester,pure6-isomer[img]/KindEditor_4.0.1/attached/image/20130704/20130704102819_7906.jpg[/img]AlexaFluor568编号：AGF1326A
Cy3-N-羟基琥珀酰亚胺酯
Cy3NHSester[img]/KindEditor_4.0.1/attached/image/20130704/20130704103024_8531.jpg[/img]AlexaFluor546NHSester
DyLight549NHSester
编号：AGF1330A
Cy3.5-N-羟基琥珀酰亚胺酯
Cy3.5NHSester[img]/KindEditor_4.0.1/attached/image/20130704/20130704103242_2437.jpg[/img]AlexaFluor594NHSester
DyLight594NHSester
编号：AGF1338A
Cy5-N-羟基琥珀酰亚胺酯
Cy5NHSester[img]/KindEditor_4.0.1/attached/image/20130704/20130704103422_4468.jpg[/img]AlexaFluor647NHSester
DyLight649NHSester
编号：AGF1345A
Cy5.5-N-羟基琥珀酰亚胺酯
Cy5.5NHSester[img]/KindEditor_4.0.1/attached/image/20130704/20130704103557_5875.jpg[/img]AlexaFluor680NHSester
DyLight680NHSester
编号：AGF1349A
Cy7-N-羟基琥珀酰亚胺酯
Cy7NHSester[img]/KindEditor_4.0.1/attached/image/20130704/20130704103713_8843.jpg[/img]编号：AGF1356A
Cy7.5-N-羟基琥珀酰亚胺酯
Cy7.5NHSester[img]/KindEditor_4.0.1/attached/image/20130704/20130704103756_5406.jpg[/img]编号：AGF1374A
磺酸基-Cy3-N-羟基琥珀酰亚胺酯
Sulfo-Cy3NHSester[img]/KindEditor_4.0.1/attached/image/20130704/20130704103830_1656.jpg[/img]AlexaFluor546
DyLight549
编号：AGF1377A
磺酸基-Cy5-N-羟基琥珀酰亚胺酯
Sulfo-Cy5NHSester[img]/KindEditor_4.0.1/attached/image/20130705/20130705095752_6656.jpg[/img]AlexaFluor647
DyLight649
编号：AGF1379A
磺酸基-Cy7-N-羟基琥珀酰亚胺酯
Sulfo-Cy7NHSester[img]/KindEditor_4.0.1/attached/image/20130704/20130704104047_6968.jpg[/img]相似系列产品：
羰基活性荧光染料
氨基类染料
巯基反应性染料
羧酸类染料
2、苏丹三 脂肪 橙红
3、苏丹四 脂肪 红
4、双缩脲 蛋白质 紫
5、龙胆紫 染色质 紫
6、碘 淀粉 蓝
7、健那绿 线粒体 绿
8、甲基绿 DNA 绿
9、吡罗红 RNA 红
10、溴麝香草酚蓝 CO2 由蓝变绿再变黄
11、重铬酸钾 酒精 酸性条件下由橙色变成灰绿
12、醋酸洋红（龙胆紫、改良苯酚品红） 染色质 红
13、台盼蓝 检验活死细胞 死细胞会被染成蓝色（不常用）
而且样品中的无水硫酸钠未变色,而做标准曲线的五个和空白对照的变为蓝色了,请高手指教,多谢!
还有,是否变蓝对测定结果有影响吗?
谢了哈
欢迎你！请下次规范发贴:)
健那绿——高中唯一一个活体染色剂。染线粒体的，染成蓝绿色
二苯胺
甲基绿派洛宁
还有一种忘了。
假期没法联系老师呵呵。
这三种的作用都是什么。
产品主要应用：点击化学(Clickchemistry)、蛋白质组学研究中的双向荧光差异凝胶电泳(2DDIGE)和实时荧光定量PCR(RealtimePCR)。
氨基类染料是包含自由氨基的活性染料，染料可与活化羧酸衍生物和其他亲电子的试剂结合。比如：氨基与EDC-活化的羧基结合。
相关产品如下：
中文名英文名产品编号分子结构Cy7.5胺Cy7.5amineAGF1350A[img]/KindEditor_4.0.1/attached/image/20130704/20130704110804_5250.jpg[/img]Cy5胺Cy5amineAGF1332A[img]/KindEditor_4.0.1/attached/image/20130704/20130704110715_7750.jpg[/img]相似系列产品：
抗体、核酸、蛋白质等生物分子标记染料
羰基活性荧光染料
巯基反应性染料
羧酸类染料
请各位大侠给予帮助！！
谢谢！！
健那绿染液是一种活体染液，实验对象必须是活细胞，健那绿可以使活细胞中的线粒体呈现蓝绿色，而细胞质接近无色。
