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Alamar Blue 阿尔玛蓝细胞增殖及毒性检测试剂 - 上海懋康生物科技有限公司
Alamar Blue 阿尔玛蓝细胞增殖及毒性检测试剂 - 上海懋康生物科技有限公司
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>Alamar Blue 阿尔玛蓝细胞增殖及毒性检测试剂
Alamar Blue 阿尔玛蓝细胞增殖及毒性检测试剂
时间:2017-04-13
作者:懋康原创
文章来源:懋康生物
Alamar Blue 阿尔玛蓝细胞增殖及毒性检测试剂
产品标签:
细胞增殖和毒性检测(Cell proliferation and cytotoxicity);MTT 噻唑兰;CCK-8;WST-1;EdU 5-乙炔基-2’-脱氧尿苷;
基本描述:
阿尔玛蓝(Alamar Blue)是一种细胞活力检测试剂,包含一种具细胞膜渗透性、无毒性且弱蓝色荧光的指示剂,即刃天青(resazurin)。自从1993年开始刃天青就作为一种值得信赖和可靠的试剂被引用。阿尔玛蓝是MTT(噻唑兰)的有效且无毒替代物。阿尔玛蓝能定量检测人、哺乳动物、细菌、真菌和支原体细胞的增殖情况,也能用于细胞因子生物活性、细胞活力分析、体外细胞毒性确定以及细胞生长监测。
工作原理:
刃天青(resazurin)是一种氧化还原(REDOX)指示剂,根据细胞代谢还原情况发生颜色变化。氧化态的刃天青呈紫蓝色且基本无荧光,其还原态产物试卤灵(resorufin)转变为粉红色且高度荧光,产生的荧光强度与呼吸活细胞数量呈正比。通过检测呼吸过程中氧化水平,阿尔玛蓝用作一种定量检测细胞活力和毒性的直接指示剂。阿尔玛蓝的颜色变化可通过普通分光光度计检测吸光度变化,检测波长为570nm,参考波长为600nm;阿尔玛蓝的荧光变化可通过荧光光度计检测,激发波长在530~560nm之间,发射波长为590nm。
产品优势:
细胞增殖与毒性检测试剂种类很多,最传统且应用广泛的有MTT噻唑兰,然MTT具有很多缺陷(比如检测时间长,对细胞毒性大,且代谢产物需要用有机溶剂溶解后才能检测等)驱使科学家不断开发和寻找更好的试剂。随后出现的有XTT,WST-1,CCK(WST-8),以及Alamar Blue阿尔玛蓝。
Alamar Blue阿尔玛蓝是最受欢迎替代MTT的检测试剂之一,优势见下:
1. 操作简单:只需添加到细胞培养物,简单孵育就可测定结果
。。。水溶性(不需清洗/固定和提取步骤);
。。。培养板可2-8°C保存且1-3天内测量结果;
2. 使用灵活
。。。适用于大多数细胞类型,包括细菌、酵母、真菌、原核生物和培养的哺乳动物细胞
。。。适用于贴壁细胞核悬浮培养细胞检测
。。。适合生色法(Colorimetric)或荧光法(Fluorometric)检测
。。。简单放大检测用于高通量分析
3. 非常安全
不同于其他检测细胞增殖的氧化还原指示剂,Alamar Blue无毒,不会干扰电子传递链,且不会影响细胞呼吸或功能。
。。。对细胞代谢的干扰最低;
。。。对细胞无毒性;
。。。对使用者无毒性;
。。。不会致癌;
。。。对环境无毒;
4. 高灵敏度:至少能检测50个细胞
5. 稳定性强
独特缓冲体系使得Alamar Blue适用于长期研究,重复检测,且与大量培养基兼容。
。。。允许连续性监测细胞
。。。酚红不会干扰结果
6. 经济实惠
不需要细胞裂解,保证细胞能够继续被培养和用于其他分析
7. 优于其他细胞活力检测试剂(MTT)
与MTT检测相比,敏感性更高且不会干扰电子链传递。
8. 应用广泛
。。。细胞增殖分析
。。。相对毒性分析
。。。细胞因子实验
订购信息:原料来自专利持有生产商有,高品质产品,现货供应,质量保证,全球已有大量的忠实粉。欢迎来电咨询,021-54736159,QQ:2971634497。
产品名称
产品编号
规格
价格(元)
货期
Alamar Blue 阿尔玛蓝细胞增殖及毒性检测试剂
MX3006-5ML
5ml (500T)
329
现货
Alamar Blue 阿尔玛蓝细胞增殖及毒性检测试剂
MX3006-10ML
10ml (1000T)
599
现货
Alamar Blue 阿尔玛蓝细胞增殖及毒性检测试剂
MX3006-50ML
50ml (5000T)
2050
现货
【产品的使用方法见说明书】。
【点击阿尔玛蓝应用示例:最佳铺板密度和孵育时间测定】。
【点击阿尔玛蓝常见应用推荐文献】
【点击阿尔玛蓝LD50测定方法】
相关产品:欢迎来电咨询,021-54736159,QQ:2971634497。【见我司懋康生物整理的不同细胞增殖检测法比较表】
产品名称
产品编号
规格
价格(元)
货期
MTT 细胞增殖及细胞毒性检测试剂盒
MX3005-500T
500T
210
现货
WST-1 细胞增殖及细胞毒性检测试剂盒
MX3007-500T
500T
450
现货
CCK-8 细胞增殖及毒性检测试剂
MX3008-1ML
1ml
(100T)
125
现货
CCK-8 细胞增殖及毒性检测试剂
MX3008-5ML
5ml
(500T)
500
现货
alamarBlue® Frequently
Asked Questions:
1. Which cells can I use for
alamarBlue tests?
2. Will
phenol red affect the measurements for alamarBlue?
3. Which
media can be used with alamarBlue?
4. Will
alamarBlue be toxic to my cells?
5. Will
microbial contaminants or carrier protein interfere with
alamarBlue measurements?
6. Why is it
necessary to optimize the cell density used for studies with alamarBlue?
7. There
seems to be a decrease in the reduction of alamarBlue if I use a long
incubation time?
8. Can I use
alamarBlue to repeatedly measure cell proliferation at intervals of days
or weeks?
9. Is it
possible to use different filters than 570 nm and 600 nm for measuring cell
growth by absorbance?
10. Is it
better to measure cell cytotoxicity using absorbance or fluorescence?
11. Can I
store my plates for measurement at a later date?
12. Will
temperature affect the fluorescence readings?
13. Can
alamarBlue be used for assays to measure lymphocyte proliferation?
14. How can I
determine the absorbance or fluorescence for the fully reduced form of
alamarBlue?
15. What are
example absorbance and fluorescence values for the oxidized and reduced forms
of alamarBlue?
— — Written/Edited by V. Shallan【版权归MKBio懋康所有】
上海懋康生物科技有限公司是一家涉足于生命科学和生物技术领域研究的试剂、仪器和实验室消耗品与实验服务工作,主要从事细胞生物学、植物学、分子生物学、免疫学、生物化学、蛋白组学。生物制药与诊断试剂研发生产等领域。 本公司秉承“以人为本,以诚为信、合同守信”的经营理念。坚持"品质保障"的原则为广大客户提供优质产品。
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alamarBlue™ Cell Viability Reagent
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HomeShop All ProductsCell Analysis ProductsCell Based AssaysCell Function Reagents and KitsalamarBlue™ Cell Viability ReagentInvitrogen™alamarBlue™ Cell Viability ReagentCatalog number: DAL1025Related applications: Cell Viability, Proliferation & FunctionCell Counting, Viability, & CryopreservationTechnical SupportCustomer ServiceInvitrogen™alamarBlue™ Cell Viability ReagentCatalog number: DAL1025Related applications: Cell Viability, Proliferation & FunctionCell Counting, Viability, & CryopreservationTechnical SupportCustomer ServiceCatalog NumberDAL1025Unit Size25 mLPrice (USD)Contact Us ›Save to listCatalog NumberDAL1100Unit Size100 mLPrice (USD)Contact Us ›Save to listCatalog NumberUnit SizePrice (USD)DAL102525 mLContact Us ›DAL1100100 mLContact Us ›Save to listProduct OverviewFiguresVideosRecommendationsRecommendationsDocumentsFAQCitations & ReferencesAdditional InformationRecommendationsalamarBlue Cell Viability Reagent is a ready-to-use resazurin-based solution that functions as a cell health indicator by using the reducing power of living cells to quantitatively measure viability. Resazurin, the active ingredient of alamarBlue reagent, is a non-toxic, cell-permeable compound that is blue in color and virtually non-fluorescent. Upon entering living cells, resazurin is reduced to resorufin, a compound that is red in color and highly fluorescent. Changes in viability can be easily detected using either an absorbance- or fluorescence-based plate reader. alamarBlue Cell Viability Reagent has broad applicability and can be used with various human and animal cell lines, bacteria, plant, and fungi.Features include:• Robust and reliable performance—results in a large, highly reproducible dynamic range • Highly sensitive reagent with a linear response—detects as few as 50 cells per well• Convenient add-and-read format—no mixing, no washing, no cell lysis• Compatible with either fluorescence- or absorbance-based instrumentation• Measures viability from many diverse cell types—including mammalian cells, bacteria, plant, and fungi Cell health can be monitored by numerous methods. Plasma membrane integrity, DNA synthesis, DNA content, enzyme activity, presence of ATP, and cellular reducing conditions are known indicators of cell viability and death. alamarBlue Cell Viability Reagent is an indigo-colored, non-toxic reagent that detects metabolically active cells and is used for the quantitative analysis of cell viability and proliferation. alamarBlue reagent has broad applicability and can be used with various human and animal cell lines, bacteria, plant, and fungi. When added to cells, alamarBlue Cell Viability Reagent is modified by the reducing environment of viable cells and turns red in color and becomes highly fluorescent. This color change and increased fluorescence can be detected using absorbance (detected at 570 and 600 nm) or fluorescence (using an excitation between 530–560 and an emission at 590 nm). To assay for viability, simply add the pre-mixed alamarBlue reagent to cells in complete media (no wash or cell lysis steps required), incubate for one to four hours, and read using either an absorbance- or fluorescence-based plate reader. If necessary, longer incubation times may be used for greater sensitivity without compromising cell health. The homogeneous, add-and-read assay format of the alamarBlue Cell Viability Reagent is fast, convenient, and readily amenable to automation and high-throughput assays. The color change and fluorescence increase resulting from cell viability changes allow the detection of alamarBlue reagent by either an absorbance- or fluorescence-based instrumentation.For Research Use Only. Not for use in diagnostic procedures.SpecificationsCell TypeBacteria, Eukaryotic Cells, Fungal CellsColorBlueDescriptionalamarBlue™ Cell Viability ReagentDetection MethodColorimetric, FluorescenceFor Use With (Application)Viability AssayFor Use With (Equipment)Spectrophotometer, Microplate ReaderProduct TypeCell Viability ReagentDye TypeResazurinEmission590 nmExcitation Wavelength Range530-560 nmFormLiquidFormat96-well plate, Cuvettes, 384-well plateIncubation Time1 to 4 hr. (Up to 24 hr. for low cell numbers)Product LinealamarBlue™Quantity25 mLShipping ConditionWet IceThroughputHigh-throughput CompatibleContents & StorageStore in refrigerator at 2°C to 8°C.FiguresDocuments & DownloadsCertificatesSearch by lot number or partial lot numberSearchRequest a CertificateFrequently asked questions (FAQs)Citations & ReferencesSearch citations by name, author, journal title or abstract textSearch
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Alamar Blue 阿尔玛蓝细胞增殖及毒性检测试剂 - 上海懋康生物科技有限公司
Alamar Blue 阿尔玛蓝细胞增殖及毒性检测试剂 - 上海懋康生物科技有限公司
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Alamar Blue 阿尔玛蓝细胞增殖及毒性检测试剂
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Alamar Blue 阿尔玛蓝细胞增殖及毒性检测试剂
目录号
MX3006-5ML
售价
329.00元
规格
5ml (500T)
运输温度
冰袋
其他名称
Alamar Blue 阿尔玛蓝;刃天青;
保存温度
CAS号
N/A
有效期
至少1年
应用
细胞增殖和毒性检测
订购数量
产品简介:
Alamar Blue 阿尔玛蓝细胞增殖及毒性检测试剂
产品标签
细胞增殖和毒性检测(Cell proliferation and cytotoxicity);MTT 噻唑兰;CCK-8;WST-1;EdU 5-乙炔基-2’-脱氧尿苷;
产品信息
产品名称
产品编号
规格
价格(元)
货期
Alamar Blue 阿尔玛蓝细胞增殖及毒性检测试剂
MX3006-5ML
5ml (500T)
329
现货
Alamar Blue 阿尔玛蓝细胞增殖及毒性检测试剂
MX3006-10ML
10ml (1000T)
599
现货
Alamar Blue 阿尔玛蓝细胞增殖及毒性检测试剂
MX3006-50ML
50ml (5000T)
2050
现货
【温馨提示】:见我司(懋康生物)整理的阿尔玛蓝最佳孵育时间和细胞密度的使用示例。
【温馨提示】:见我司(懋康生物)整理的阿尔玛蓝测定LD50的方法。
【温馨提示】:见我司(懋康生物)整理的阿尔玛蓝常见问题(FAQs)。
【温馨提示】:见我司(懋康生物)整理的阿尔玛蓝常见应用推荐文献。
产品描述
阿尔玛蓝(Alamar Blue)是一种细胞活力检测试剂,包含一种具细胞膜渗透性、无毒性且弱蓝色荧光的指示剂,即刃天青(resazurin)。自从1993年开始刃天青就作为一种值得信赖和可靠的试剂被引用。阿尔玛蓝是MTT(噻唑兰)的有效且无毒替代物。阿尔玛蓝能定量检测人、哺乳动物、细菌、真菌和支原体细胞的增殖情况,也能用于细胞因子生物活性、细胞活力分析、体外细胞毒性确定以及细胞生长监测。
阿尔玛蓝的工作原理在于:刃天青是一种氧化还原(REDOX)指示剂,根据细胞代谢还原情况发生颜色变化。氧化态的刃天青呈紫蓝色且基本无荧光,其还原态产物试卤灵(resorufin)转变为粉红色且高度荧光,产生的荧光强度与呼吸活细胞数量呈正比。通过检测呼吸过程中氧化水平,阿尔玛蓝用作一种定量检测细胞活力和毒性的直接指示剂。阿尔玛蓝的颜色变化可通过普通分光光度计检测吸光度变化,检测波长为570nm,参考波长为600nm;阿尔玛蓝的荧光变化可通过荧光光度计检测,激发波长在530~560nm之间,发射波长为590nm。
与台盼蓝、TTC、MTT、MTS等分析法相比,阿尔玛蓝为单一试剂,可连续快速的检测细胞的增殖情况。阿尔玛蓝对细胞无毒无害,不会干扰电子传递链,不会干扰细胞呼吸或功能,也不会影响细胞的抗体合成与分泌等活性,因此适用于对同一批细胞的增殖情况进行连续观察和更进一步的观察,具有操作简便和几乎不干扰正常代谢的特征。
保存与运输方法
保存:4℃避光保存,至少一年有效。
运输:冰袋运输。
注意事项
1. 还原态的Alamar Blue在水或水溶性缓冲液如PBS中很不稳定,但在培养基内非常稳定。为了确定特定实验还原态Alamar Blue的吸光度/荧光值,需准备100%还原态Alamar Blue试剂:将Alamar Blue与细胞培养基按照1:10的比例混合,高压灭菌15min即可。
2. 适宜的细胞密度能够增加检测灵敏度。对于96孔板,建议每孔接种100μl细胞,细胞浓度范围:贴壁细胞为100-10,000个/孔,悬浮细胞在2,000-50,000个/孔,以培养基为空白对照。对于384孔板,细胞浓度和接种量均减半。
3. 影响细胞对Alamar Blue反应的两大变量为孵育时间和铺板密度,建议实验前需先摸索优化这两个因素。细胞密度过高或孵育时间过长都会导致继发性还原反应,红色产物停止增加并开始衰减,导致吸光度/荧光水平明显下跌并伴随红色消失。
4. 微生物污染会还原Alamar Blue。因此对被污染细胞使用Alamar Blue检测会引起错误结果。
5. Alamar Blue可以用分光光度计或荧光计检测,但荧光灵敏度高,实验误差小,推荐荧光检测。
6. 为了您的安全和健康,请穿实验服并戴一次性手套操作。
使用方法
一、标准曲线制作(最佳孵育时间和细胞密度的测定)
1. 收集对数生长期的细胞并进行细胞计数。每孔接种100μl细胞悬液,按比例依次用培养基等比稀释成浓度梯度,一般做3-5个浓度梯度,每个浓度做3-6个复孔。注意要设置阴性对照(细胞培养基中不加入细胞)和阳性对照(只加100μl 100%还原态Alamar Blue,不含细胞)。
2. 每孔加入10μlAlamar Blue,阳性对照孔加入10μl无菌超纯水。
3. 将平板放回细胞培养箱孵育。接下来的6-8h内每隔1h取出平板并测定荧光/吸光值。建议平板孵育过夜并测定第24h的荧光/吸光值,得到以下两组数据:
a)在任何选定的孵育时间内,与细胞数量相关的细胞密度对Alamar Blue还原水平通过线性反应来确定;
b)在任何选定的细胞密度上,测定随着对照细胞所需时间将Alamar Blue从氧化态(蓝色)转化为完全还原态(红色)来确定所需的孵育时间;
4.推荐用荧光分光光度计检测,激发光波长在530-560 nm之间,发射光波长为590 nm。或者用普通分光光度计在570nm测定吸光值,参考波长600nm。也可用570/630nm和540/600nm替代。
5.计算在每个特定细胞密度或孵育时间上Alamar Blue的还原率。
a)公式1:通过吸光值来计算还原率
Alamar Blue还原率(%)=[(E600×A570)-(E570×A600)]/[(E570′×C600)-(E600′×C570)] ×100
E600=氧化态Alamar Blue在600nm波长的消光系数;
E570=氧化型Alamar Blue在570nm波长的消光系数;
A600=检测孔在600nm波长的吸光值;
A570=检测孔在570nm波长的吸光值;
E570′=还原态Alamar Blue在570nm波长的消光系数;
E600′=还原态Alamar Blue在600nm波长的消光系数;
C570=阴性对照孔在570nm波长的吸光值(不加细胞的培养基+Alamar Blue);
C600=阴性对照孔在600nm波长的吸光值(不加细胞的培养基+Alamar Blue);
表1. 阿尔玛蓝(Alamar Blue)在不同波长下的消光系数
波长(nm)
还原态 Alamar Blue
氧化态Alamar Blue
540
104395
47619
570
155677
80586
600
14652
117216
630
5494
34798
b)公式2:通过荧光值来计算还原率
Alamar Blue还原率(%)=(实验组FI 590-阴性对照组FI 590)/(100%还原态Alamar Blue阳性对照FI 590-阴性对照组FI 590)×100
FI 590=590nm发射波长(560nm激发波长)下的荧光强度;
6. 绘制在每个特定细胞密度或孵育时间Alamar Blue的还原率。
a)对确定最佳细胞密度的实验,以细胞密度的对数(log)为x坐标,Alamar Blue还原率为纵坐标;
b)对确定最佳孵育时间的实验,以孵育时间为x坐标,Alamar Blue还原率为纵坐标;
c)根据以上曲线图来确定最佳细胞密度或孵育时间。
二、细胞毒性和细胞增殖检测
1.收集对数生长期的细胞并进行细胞计数。调整细胞数为1×104个/ml(建议细胞密度),每孔接种100μl细胞悬液。最佳细胞密度需根据细胞类型决定。
2.细胞铺板,并加入待检测药物。为了确定药物对细胞生长影响,需设置合适的对照组,包括刺激和无刺激细胞组。
3.振荡混匀,放入细胞培养箱内孵育一段时间。
4.无菌条件下每孔加入10μlAlamar Blue。阳性对照孔中加入10μl无菌的超纯水。
5.放入细胞培养箱内孵育4-8h,最佳孵育时间取决于细胞类型。
6.使用光度计测定细胞毒性或增殖情况。
7.推荐用荧光分光光度计检测,激发光波长在530-560 nm之间,发射光波长为590 nm。或者用普通分光光度计在570nm测定吸光值,参考波长600nm。使用培养基为空白对照。
8.计算细胞毒性和增殖实验下对照细胞和处理细胞间Alamar Blue还原度的变化百分比。
a)公式3:通过吸光值来计算还原度的变化百分比
Alamar Blue还原度变化百分比(%)=[(E600×A570)-(E570×A600)]/[( E600×P570)-(E570×P600)] ×100
E600=氧化态Alamar Blue在600nm波长的消光系数;
E570=氧化态Alamar Blue在570nm波长的消光系数;
A600=检测孔在600nm波长的吸光值;
A570=检测孔在570nm波长的吸光值;
P570=阳性对照孔在570nm波长的吸光值(不含待测药物的细胞+ Alamar Blue)
P600=阳性对照孔在600nm波长的吸光值(不含待测药物的细胞+ Alamar Blue)
表1. 阿尔玛蓝(Alamar Blue)在不同波长下的消光系数
波长(nm)
还原态 Alamar Blue
氧化态Alamar Blue
540
104395
47619
570
155677
80586
600
14652
117216
630
5494
34798
结果判断:假如Alamar Blue还原度变化百分比(%)=62%,说明相对于对照孔,检测孔Alamar Blue的还原变化值为62%,换句话而言,相对于对照孔,38%的检测孔细胞生长受到抑制。
b)公式4:通过荧光值来计算还原度的变化百分比
Alamar Blue还原度变化百分比(%)=实验组FI 590/阴性对照组FI 590)×100
FI 590=590nm发射波长(560nm激发波长)下的荧光强度;
结果判断:假如Alamar Blue还原度变化百分比(%)=25%,说明相对于对照孔,检测孔Alamar Blue的还原变化值为25%,换句话而言,相对于对照孔,75%的检测孔细胞生长受到抑制。
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文献引用
Wang Lixuan et al. Fabrication of Injectable, Porous Hyaluronic Acid Hydrogel Based on an In-Situ Bubble-Forming Hydrogel Entrapment Process. Polymers 2020, 12(5), 1138; https://doi.org/10.3390/polym12051138
Alamar Blue was bought from Maokang Biotechnology Co., Ltd., (Shanghai, China).
To detect cytotoxicity and cell proliferation, the rBMSC suspension was added to the wells from the top of the scaffold, and culturing took place in a cell incubator at 37 °C for 1, 4, or 7 days. After the end of the culturing, we added Alamar blue reagent to the well plate and continued incubating in the cell incubator for 2–8 h until the color of the culture medium changed from blue to pink. Finally, the fluorescence intensity which is directly proportional to the number of viable cells was measured by a fluorescence photometer (SAFIRE, Tecan Co., Mannedorf, Switzerland).
Xiangyun Niu et al. Effects of Pinus massoniana pollen polysaccharides on intestinal microenvironment and colitis in mice. Food Funct., 2021,12, 252-266 https://doi.org/10.1039/D0FO02190C
After 20μl of Alamar Blue™ solution (Maokang Biotechnology, China) was added to each well, the cells were then continuously cultured for 5–24 h.
— — Written/Edited by V. Shallan【版权归MKBio懋康所有】
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alamarBlue-新一代细胞活性指示剂 - 知乎
alamarBlue-新一代细胞活性指示剂 - 知乎切换模式写文章登录/注册alamarBlue-新一代细胞活性指示剂欣博盛生物主要提供抗体,蛋白,ELISA试剂盒,核苷酸,mRNA等AlamarBlue®——简便、快速、可靠的细胞活力和增殖分析试剂AlamarBlue® 是一种快速、灵敏的细胞增殖和细胞毒性检测试剂,适用于人、动物细胞、细菌和真菌等多种研究对象,可用于贴壁细胞和非贴壁细胞的检测。作为一种无毒、即用型的试剂,alamarBlue®为细胞增殖分析提供了一种简便、快速、安全、经济、可靠的方法。 AlamarBlue® 订购信息供应商目录号名称名称使用说明AbD SerotecBUF012AalamarBlue®25 ml2500 wells /96-well plateAbD SerotecBUF012BalamarBlue®100 ml10000 wells /96-well plateAlamarBlue® 的特点与优势● 使用简单方便:即用型的单组分试剂,只需加入培养基即可检测细胞的增殖状况;产生的还原产物是水溶性的,与其他常用的细胞增殖检测方法相比,不需要固定、洗涤、提取的步骤● 灵活:可通过分光光度计或荧光光度计检测● 性价比高:可用于高通量分析● 安全:对细胞无毒、无害,不影响细胞代谢、细胞因子分泌、抗体合成等,对人体和环境也无毒无害● 高灵敏:最低可检测50个细胞● 稳定:适用于对细胞增殖状况的连续观察● 经济:无需裂解细胞,细胞可继续培养或用于其他实验● 可靠:文献引用率高,Pubmed有上千篇文献的引用AlamarBlue® 的应用● 监测细胞的生长增殖状态、研究细胞周期、细胞凋亡● 监测生长因子活性,评估生长因子对细胞增殖的影响● 治疗性药物尤其是肿瘤治疗药物的开发● 环境污染物质的评估,细胞毒分析● 抗生素效价评估AlamarBlue® 的原理细胞增殖过程中,细胞体内的环境由氧化环境变化成还原环境,呼吸链中的NADPH/NADP, FADH/FAD, FMNH/FMN和NADH/NAD的比值升高。alamarBlue®的活性成分resazurin (刃天青)是一种无毒、可透膜的蓝色染料,有微弱的荧光性,它作为一种氧化还原指示剂,被细胞内吞后,在细胞质中被以上这些代谢中间体还原,其还原产物resorufin呈现出粉红色并有很强的荧光性,最后可用普通分光光度计或荧光光度计进行检测,吸光度和荧光强度与活性细胞数成正比,因而可作为细胞增殖和细胞毒性定量检测的一个理想的指示器。AlamarBlue® 的操作流程以及结果计算操作非常简单、容易:在培养的细胞中加入alamarBlue® , 于37°C 孵育1-4小时,然后读取荧光值或吸光值(荧光检测为 530-560 nm 激发,发射光波长为590 nm ,或者在 570 nm 和 600 nm用分光光度计测定光吸收值)。使用alamarBlue® calculator 进行结果计算AbD Serotec公司专门提供在线的alamarBlue® calculators 软件,通过该软件可直接将原始的吸光值或荧光值转换为定量的结果,并可通过Excel文件格式导出。AlamarBlue® Colorimetric Calculator AlamarBlue® Fluorometric CalculatorAlamarBlue® 的保存● 室温下可保存12个月,2-8℃下可延长保质期到20个月● 该产品不得稀释储存存● 此产品对光敏感,须避光保存 注意事项alamarBlue®使用的pH范围是6.8 - 7.4。如需购买该产品,请联系AbD Serotec代理商欣博盛生物本文来源:alamarBlue-新一代细胞活性指示剂 - Bio-rad - 深圳欣博盛生物科技有限公司,转载请说明出处~发布于 2021-12-13 10:40计算药物化学细胞肿瘤细胞赞同 4添加评论分享喜欢收藏申请
alamarBlue Assays for Cell Viability | Thermo Fisher Scientific - PH
alamarBlue Assays for Cell Viability | Thermo Fisher Scientific - PH
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Home›Life Sciences›Cell Analysis›Fluorescence Microplate Assays›Microplate Assays for Cell Viability›alamarBlue Assays for Cell Viability alamarBlue Assays for Cell ViabilitySee Navigation‹Microplate Assays for Cell ViabilityalamarBlue Assays for Cell Viability
›PrestoBlue Assays for Cell Viability
›CyQUANT Cytotoxicity Assays
›CyQUANT XTT and MTT Assays for Cell Viability
›alamarBlue HS and alamarBlue Cell Viability Reagents are ready-to-use, non-toxic, resazurin-based solutions that function as cell health indicators to quantitatively measure viability. alamarBlue uses the natural reducing power of living cells to convert resazurin to fluorescent resorufin. alamarBlue HS (High Sensitivity) Cell Viability Reagent is a superior version of alamarBlue that retains all the key alamarBlue reagent characteristics, but also provides much higher sensitivity.alamarBlue HS and alamarBlue are recommended in cases of extended viability studies or when using a high cell density.On this page:Selection guideWhat are alamarBlue assays?How do I use alamarBlue assays?alamarBlue HS has improved performanceSee alsoalamarBlue Assays for Cell Viability protocolRelated content:Cell proliferation assaysCell viability assaysMicroplate assays for cell viability
Selection guide
alamarBlue HS (high sensitivity) Cell Viability ReagentalamarBlue Cell Viability ReagentUseMicroplate assay recommended in cases of extended viability studies or when using a high cell densityApplicable to a wide variety of mammalian cells, bacteria, plants, and fungiMechanism of detectionResazurin is converted to fluorescent resorufinSensitivity Detect as few as 20 cells per well Detect as few as 50 cells per wellReagent purityPurifiedVarying concentrations of resorufin contamination depending on sources of material and manufacturing conditionsBackground fluorescenceLow>50% reduction in background fluorescenceModerateIncreased background fluorescence due to presence of resorufin contaminationSignal-to-backgroundHigh>100% ratio increaseModerateReduced signal-to-background due to presence of resorufin contaminationAssay signal windowHigh>10-fold increase as compared to the non-HS assayModerateReduced dynamic range due to the presence of resorufin contaminationCat. No.A50100DAL1025alamarBlue HS reagent is recommended in cases of extended viability studies or when using a high cell density, whereas PrestoBlue HS reagent is recommended for quick viability determination (10-minute incubation).Learn more about PrestoBlue and PrestoBlue HS
What are alamarBlue assays?
Highly referenced for cytotoxicity and viability assays, alamarBlue cell viability assay has been used for years in biological and environmental studies (1). Furthermore, analysis of cell proliferation and cytotoxicity is a vital step in evaluating cellular health and in the drug discovery process. alamarBlue is a proven cell viability indicator that uses the natural reducing power of live cells to convert resazurin to the fluorescent molecule, resorufin. Analysis can be evaluated quantitatively on an absorbance- or fluorescence-based microplate reader; while qualitative analysis can be evaluated by the visual color change of the solution which is indicative of metabolically active cells.The active ingredient of alamarBlue (resazurin) is a non-toxic (Figure 1), cell permeable compound that is blue in color and virtually non-fluorescent. Upon entering cells, resazurin is reduced to resorufin, which produces a very bright red fluorescence (Figure 2). Viable cells continuously convert resazurin to resorufin, thereby generating a quantitative measure of viability—and cytotoxicity. Click image to enlargeFigure 1. Demonstrated Non-Toxicity of the alamarBlue HS Cell Viability Reagent. U2OS Cells were treated with and without alamarBlue HS for 4 hours. CyQUANT Direct reagent was added to both populations following manufacturer’s instructions. Based on the CyQUANT Direct fluorescence measurements there were no significant differences in fluorescence measurements suggesting alamarBlue HS does not significantly contribute to cellular toxicity. Additionally, when exposed to varying concentrations of gambogic acid, the U2OS cells pre-treated with alamarBlue HS displayed similar IC50 values as the control cells.Figure 2. Metabolically active cells convert resazurin to resorufin, a red-fluorescent indicator. Resazurin is a non-fluorescent indicator dye that undergoes chemical reduction to bright red-fluorescent resorufin in metabolically active cells. The amount of fluorescence produced is proportional to the number of living cells. In alamarBlue HS, the resazurin is purified and provides superior performance compared to the standard alamarBlue reagent which has known contamination issues that lead to sub-optimal performance.alamarBlue has a proven track record as an indicator of cell health and its nontoxic nature permits long-term exposure of cells without negative impact; cells grown in the presence of alamarBlue were found to produce similar numbers of viable cells as control cells, as determined by flow cytometric analysis of CD44, CD45RB, and CD4 antigens.The alamarBlue HS reagent refers to the fact that the alamarBlue HS reagent contains purified resazurin, devoid of any contaminating resorufin which can negatively impact reagent performance. The alamarBlue HS reagent provides a >50% reduction in background signal and >100% signal-to-background ratio increase compared to the standard alamarBlue reagent, while retaining all the key characteristics that make alamarBlue a highly-referenced cell viability and cytotoxicity reagent.TOP
How do I use alamarBlue assays?
alamarBlue HS and alamarBlue cell viability reagents are easy add-and-read assays (Figure 3). Simply add alamarBlue HS or alamarBlue to your cells, incubate for 1–4 hours, and read the fluorescence or absorbance. The amount of fluorescence or absorbance is proportional to the number of living cells and corresponds to the cell’s metabolic activity. Damaged and non-viable cells have lower innate metabolic activity and thus generate a proportionally lower signal than healthy cells.alamarBlue assays are compatible with multiple instrument platforms. After incubation with alamarBlue HS or alamarBlue, your samples can readily be measured on fluorescence and absorbance instrumentation. For fluorescence, simply set up your plate reader or fluorescence spectrophotometer using 560/590 nm (excitation/emission) filter settings. Alternatively, the absorbance of alamarBlue HS and alamarBlue can be read on a UV-Vis spectrophotometer at 570 nm.Finally, analyze results by plotting fluorescence intensity (or absorbance) versus compound concentration. While results are linear and quantitative for both fluorescence and absorbance, fluorescence readings provide higher sensitivity.Figure 3. How alamarBlue assays work.
alamarBlue assays do not require cell lysis
alamarBlue HS easily enters into live cells, eliminating the need to lyse or further process cells using fixation and DNA denaturation techniques. The dye is stable in cell culture media, including media containing phenol red. These characteristics allow you to:Continuously monitor the effects of drugs in a dose response assay or when optimizing drug concentrationsPerform other functional assays after cell viability measurements with alamarBlue HS or alamarBlue are completeTOP
alamarBlue HS has improved performance
As a result of the synthesis and manufacturing processes, all resazurin-based reagents contain a detectable amount of highly fluorescent resorufin contamination. The amount of the resorufin contamination can vary greatly between sources of the material and manufacturing conditions. The varying amounts of resorufin contribute to differences in detectable background fluorescence. Additionally, the contaminating resorufin and the resulting higher background signal significantly reduces the signal-to-background ratio and dynamic range of the assay.To improve the performance of the resazurin-based reagents, an innovative process was developed and, on implementation, removes the contaminating resorufin resulting in purified resazurin. This purified resazurin was used in the standard formulation creating the multiple component alamarBlue HS Cell Viability Reagent.
alamarBlue HS has higher sensitivity compared to alamarBlue
The purified resazurin used for alamarBlue HS reagent results in a >50% decrease in background fluorescence and a >100% increase in the signal-to-background ratio (Figures 4 and 5). Click image to enlargeFigure 4. Significantly lower background fluorescence observed when comparing alamarBlue HS vs alamarBlue. The purified resazurin used for the alamarBlue HS formulation results in a cell viability reagent displaying a background fluorescence reduction in suspension (Ramos), primary cells (HCASMC), and adherent (U2OS) cells.Figure 5. Performance improvement in signal to background ratio with alamarBlue HS as compared to alamarBlue. Improved cell viability signal to background ratios were displayed with HCASMC (primary cells), Ramos (suspension cells), and U2OS (adherent cells) when using the alamarBlue HS cell viability reagent which contains purified resazurin as compared to the standard alamarBlue formulation.
alamarBlue HS has a larger assay window compared to alamarBlue
The alamarBlue HS reagent displays an extended viability detection time window for a wide variety of organisms and thus can be used with various human and animal cell lines, bacteria, plant, and fungi, resulting in quantitative measurement of cell viability and proliferation (Figure 6).Figure 6. Much larger dynamic range observed with the alamarBlue HS Cell Viability Reagent which contains purified resazurin as compared to the standard alamarBlue formulation. Various concentrations of gambogic acid were added to A549 cells and the changes in viability were detected. The results demonstrate a significant improvement in the dynamic range of the assay when using alamarBlue HS.Also see poster: New and improved cellular health evaluation of 2D and 3D cellular models using microplate reader assaysTOPExpertly detect fluorescence with Thermo Scientific plate readersHigh-sensitivity fluorescence detection for 96–1,536 samples can be quickly performed on Fluoroskan or Fluoroskan FL Microplate Fluorometer or Varioskan LUX Multimode Microplate Reader using Invitrogen reagents for optimal detection. Take advantage of automatic dynamic range selection to get optimal gain settings for each individual well and automation capabilities for even higher throughput.Compare readers
Citations
Rampersad SN (2012). Sensors. 12(9):12347-12360. PMID: 23112716
Resources
Cell Analysis Learning Center—Find educational resources such as application notes, webinars, videos, articles, and more that cover the use of many of our reagents and kits for cell analysis.Fluorescence SpectraViewer—Online tool for visualization of the excitation and emission of fluorescent reagents; allows for checking spectral compatibility for multiple fluorophores.Application notes Application note: Evaluation of the toxic effects of PFOS in hESCs differentiating into cardiomyocytes Application note: Monitoring cell health with alamarBlue and PrestoBlue reagents using the Varioskan LUX Multimode Microplate ReaderBioProbes articleAn Absorbance-Based Assay for Cell Health and ProliferationPoster New and improved cellular health evaluation of 2D and 3D cellular models using microplate reader assays
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Cell Analysis Support Center—Find technical information, tips and tricks, and answers to everyday problems.5 Steps resources5 Steps to Publication-Quality Fixed Cell Imaging5 Steps to Live-Cell ImagingSee all 5 Steps WorkflowsFor Research Use Only. Not for use in diagnostic procedures.
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