CFDA (5(6)-Carboxyfluorescein diacetate) 活细胞示踪探针


描述

CFDA (5(6)-Carboxyfluorescein diacetate) 活细胞示踪探针

产品关键词:

5(6)-CFDA;Fluorescein diacetate (FDA) 二乙酸荧光素;;Calcein AM钙黄绿素;CDCFDA; CAS NO:124387-19-5;

产品信息

产品名称 产品编号 规格 价格(元)
CFDA (5(6)-Carboxyfluorescein diacetate) MX4104-25MG 25mg 452
CFDA (5(6)-Carboxyfluorescein diacetate) MX4104-100MG 100mg 992

产品描述

CFDA(5(6)-Carboxyfluorescein diacetate,CAS:124387-19-5),一种细胞膜渗透性的酯酶底物,用作一种活细胞探针,不仅能用来测定激活其荧光所需的酶活性,也能用来检测维持荧光产物保留在胞内的细胞膜完整性。一旦被胞内酯酶水解后,代谢产生羧基荧光素(Carboxyfluorescein, CF)。与荧光素相比,羧基荧光素含有更多的负电荷,从而能更好的保留在胞内。

产品特性

1) CAS NO:124387-19-5

2) 化学名:Spiro(isobenzofuran-1(3H),9′-(9H)xanthene)-ar-carboxylic acid, 3′,6′-bis(acetyloxy)-3-oxo-

3) 同义名:5(6)-CFDA; 5(6)-CFDA, mixed isomers;5-(and-6)-Carboxyfluorescein Diacetate, mixed isomers; 5(6)-Carboxy-di-O-acetylfluorescein; 5(6)-羧基荧光素二乙酸酯;

4) 分子式:C25H16O9

5) 分子量:460.39 g/mol

6) 外观:浅黄色至深黄色粉末

7) 纯度:≥90%(HPLC)

8) EX/Em:492/517nm (0.1 M Tris pH 8.0 (esterase))

9) 溶解性:溶于DMSO

10)化学结构式:

保存与运输方法

保存:2-8℃避光干燥保存,长期不用可置于-20℃避光干燥保存,至少2年有效。

运输:室温运输。

常见应用

CFDA(5(6)-Carboxyfluorescein diacetate,CAS:124387-19-5)常用来检测细菌活性和代谢活性。多与细胞核染料碘化丙啶(PI)或DAPI联合使用,以下列出此探针的一些应用文献,仅供交流学习用。

另外有资料显示,革兰氏阴性菌用CFDA染色所得的荧光强度会低于革兰氏阳性菌,原因在于,革兰氏阴性菌的外膜结构会在一定程度上阻挡CFDA进入细胞。对于革兰氏阴性菌染色,可使用添加含EDTA的染色缓冲液(比如0.1M-Phosphate buffer (pH8.5), 5%(w/v) NaCl, 0.5 mM EDTA)或戊二醛(参考文献:Morono Y et al. Application of glutaraldehyde for the staining of esterase-active cells with carboxyfluorescein diacetate. Biotechnol Lett. 2004 Mar;26(5):379-83.)

文献1)Yamaguchi N et al. Rapid monitoring of bacteria in dialysis fluids by fluorescent vital staining and microcolony methods. Nephrol Dial Transplant. 2007 Feb;22(2):612-6. Epub 2006 Nov 22.

检测背景:CFDA是一种比较流行的、灵敏的荧光底物用于预估活微生物(细菌、酵母或真菌)的酯酶活性。细胞质内的酯酶活性是微生物生理活性的一种指标。通过CFDA-DAPI双染色方法同时确定总细菌数和具有生理活性的细菌数(具有酯酶活性)。

检测对象:500ml透析液样本的细菌细胞

检测方法:Bacterial cells in 500 ml of dialysis fluid samples were vacuum-filtered onto polycarbonate black filters and then incubated in staining buffer [0.1 mol/l phosphate buffer (pH8.5), 5% (w/v) NaCl, 0.5 mmol/l EDTA (ethylenediaminetetraacetic acid)] containing DAPI (final concentration, 1 μg/ml) and CFDA (final concentration, 150 μg/ml) in a filtration funnel for 5 min at room temperature (∼25°C). The staining buffer in the funnel was then removed by vacuum filtration. The filters were mounted on glass slides with immersion oil and fluorescing cells were counted at 1000 × magnification using an E-400 epifluorescence microscope equipped with a 100 W Hg lamp.

检测结果:


Fig 1.Bacterial cells in dialysis fluid. (A) Total bacterial cells stained with 4′,6-diamidino-2-phenylindole (DAPI) and observed under ultraviolet excitation. (B) Esterase-active cells stained with carboxyfluorescein diacetate (CFDA) and observed under blue excitation. (C) Microcolonies stained with SYBR Gold and observed under blue excitation.

文献2)Majeed M et al. Rapid assessment of viable but non-culturable Bacillus coagulans MTCC 5856 in commercial formulations using Flow cytometry. PLoS One. 2018 Feb 23;13(2):e0192836.

检测对象:B. coagulans MTCC 5856 spores

检测方法:Double staining with CFDA and PI were carried out by incubating the spore suspension (107/ml) with 50μM CFDA at 37°C for 60 minutes in and 10μM PI in the last 10 min. The time for CFDA staining was standardized earlier at different times and 60 minutes was found to be optimum.

检测结果:

Fig 2. Flow cytometric analysis of B coagulans MTCC 5856 spores.10 mg of dry spores were suspended in 1ml of PBS and activated for 30 min at 75°C, followed by fluorescent staining as described in the methods section. Data represents mean± standard deviation of at least 3 independent experiments. A: Gating strategy for SYTOBC ™ staining- The microbeads and the stained spores are gated separately based on unstained control B: Gating strategy for CFDA and PI staining. The dot plot represents a double staining for both CFDA and PI. The quadrant 4(Q4) represents live cells, Q1- dead cells and Q2 damaged cells. The double positive cells were not considered for enumerating the live spores. C: Representative dot plot for the FCM analysis of Live and Dead spores D: Table showing the calculation of viable spores.

注意事项

1) 本品保存过程中远离氧化剂。

2) 为了您的安全和健康,请穿实验服并戴一次性手套操作。

相关产品

货号 名称 规格
MX3009-25MG CFDA, SE (CFSE)细胞增殖示踪荧光探针 25mg
MX3010-500T CFDA SE Cell Proliferation and Cell Tracking Kit 500T
MX3011-50UG Calcein, AM, Ultra Pure Grade钙黄绿素(绿色) 50μg
MX3012-500T Calcein AM/PI Double Stain Kit活细胞/死细胞双染试剂盒 500T
MX4101-1G Fluorescein diacetate (FDA)二乙酸荧光素 1g
MX4102-25MG CDCFDA, SE活细胞示踪探针 25mg
MX4103-100MG CDCFDA (5(6)-Carboxy-2’,7’-dichlorofluorescein diacetate)活细胞示踪探针 100mg
MX4104-25MG CFDA (5(6)-Carboxyfluorescein diacetate)活细胞示踪探针 25mg

CFDA (5(6)-Carboxyfluorescein diacetate) 活细胞示踪探针


描述

CFDA (5(6)-Carboxyfluorescein diacetate) 活细胞示踪探针

产品关键词:

5(6)-CFDA;Fluorescein diacetate (FDA) 二乙酸荧光素;;Calcein AM钙黄绿素;CDCFDA; CAS NO:124387-19-5;

产品信息

产品名称 产品编号 规格 价格(元)
CFDA (5(6)-Carboxyfluorescein diacetate) MX4104-25MG 25mg 452
CFDA (5(6)-Carboxyfluorescein diacetate) MX4104-100MG 100mg 992

产品描述

CFDA(5(6)-Carboxyfluorescein diacetate,CAS:124387-19-5),一种细胞膜渗透性的酯酶底物,用作一种活细胞探针,不仅能用来测定激活其荧光所需的酶活性,也能用来检测维持荧光产物保留在胞内的细胞膜完整性。一旦被胞内酯酶水解后,代谢产生羧基荧光素(Carboxyfluorescein, CF)。与荧光素相比,羧基荧光素含有更多的负电荷,从而能更好的保留在胞内。

产品特性

1) CAS NO:124387-19-5

2) 化学名:Spiro(isobenzofuran-1(3H),9′-(9H)xanthene)-ar-carboxylic acid, 3′,6′-bis(acetyloxy)-3-oxo-

3) 同义名:5(6)-CFDA; 5(6)-CFDA, mixed isomers;5-(and-6)-Carboxyfluorescein Diacetate, mixed isomers; 5(6)-Carboxy-di-O-acetylfluorescein; 5(6)-羧基荧光素二乙酸酯;

4) 分子式:C25H16O9

5) 分子量:460.39 g/mol

6) 外观:浅黄色至深黄色粉末

7) 纯度:≥90%(HPLC)

8) EX/Em:492/517nm (0.1 M Tris pH 8.0 (esterase))

9) 溶解性:溶于DMSO

10)化学结构式:

保存与运输方法

保存:2-8℃避光干燥保存,长期不用可置于-20℃避光干燥保存,至少2年有效。

运输:室温运输。

常见应用

CFDA(5(6)-Carboxyfluorescein diacetate,CAS:124387-19-5)常用来检测细菌活性和代谢活性。多与细胞核染料碘化丙啶(PI)或DAPI联合使用,以下列出此探针的一些应用文献,仅供交流学习用。

另外有资料显示,革兰氏阴性菌用CFDA染色所得的荧光强度会低于革兰氏阳性菌,原因在于,革兰氏阴性菌的外膜结构会在一定程度上阻挡CFDA进入细胞。对于革兰氏阴性菌染色,可使用添加含EDTA的染色缓冲液(比如0.1M-Phosphate buffer (pH8.5), 5%(w/v) NaCl, 0.5 mM EDTA)或戊二醛(参考文献:Morono Y et al. Application of glutaraldehyde for the staining of esterase-active cells with carboxyfluorescein diacetate. Biotechnol Lett. 2004 Mar;26(5):379-83.)

文献1)Yamaguchi N et al. Rapid monitoring of bacteria in dialysis fluids by fluorescent vital staining and microcolony methods. Nephrol Dial Transplant. 2007 Feb;22(2):612-6. Epub 2006 Nov 22.

检测背景:CFDA是一种比较流行的、灵敏的荧光底物用于预估活微生物(细菌、酵母或真菌)的酯酶活性。细胞质内的酯酶活性是微生物生理活性的一种指标。通过CFDA-DAPI双染色方法同时确定总细菌数和具有生理活性的细菌数(具有酯酶活性)。

检测对象:500ml透析液样本的细菌细胞

检测方法:Bacterial cells in 500 ml of dialysis fluid samples were vacuum-filtered onto polycarbonate black filters and then incubated in staining buffer [0.1 mol/l phosphate buffer (pH8.5), 5% (w/v) NaCl, 0.5 mmol/l EDTA (ethylenediaminetetraacetic acid)] containing DAPI (final concentration, 1 μg/ml) and CFDA (final concentration, 150 μg/ml) in a filtration funnel for 5 min at room temperature (∼25°C). The staining buffer in the funnel was then removed by vacuum filtration. The filters were mounted on glass slides with immersion oil and fluorescing cells were counted at 1000 × magnification using an E-400 epifluorescence microscope equipped with a 100 W Hg lamp.

检测结果:


Fig 1.Bacterial cells in dialysis fluid. (A) Total bacterial cells stained with 4′,6-diamidino-2-phenylindole (DAPI) and observed under ultraviolet excitation. (B) Esterase-active cells stained with carboxyfluorescein diacetate (CFDA) and observed under blue excitation. (C) Microcolonies stained with SYBR Gold and observed under blue excitation.

文献2)Majeed M et al. Rapid assessment of viable but non-culturable Bacillus coagulans MTCC 5856 in commercial formulations using Flow cytometry. PLoS One. 2018 Feb 23;13(2):e0192836.

检测对象:B. coagulans MTCC 5856 spores

检测方法:Double staining with CFDA and PI were carried out by incubating the spore suspension (107/ml) with 50μM CFDA at 37°C for 60 minutes in and 10μM PI in the last 10 min. The time for CFDA staining was standardized earlier at different times and 60 minutes was found to be optimum.

检测结果:

Fig 2. Flow cytometric analysis of B coagulans MTCC 5856 spores.10 mg of dry spores were suspended in 1ml of PBS and activated for 30 min at 75°C, followed by fluorescent staining as described in the methods section. Data represents mean± standard deviation of at least 3 independent experiments. A: Gating strategy for SYTOBC ™ staining- The microbeads and the stained spores are gated separately based on unstained control B: Gating strategy for CFDA and PI staining. The dot plot represents a double staining for both CFDA and PI. The quadrant 4(Q4) represents live cells, Q1- dead cells and Q2 damaged cells. The double positive cells were not considered for enumerating the live spores. C: Representative dot plot for the FCM analysis of Live and Dead spores D: Table showing the calculation of viable spores.

注意事项

1) 本品保存过程中远离氧化剂。

2) 为了您的安全和健康,请穿实验服并戴一次性手套操作。

相关产品

货号 名称 规格
MX3009-25MG CFDA, SE (CFSE)细胞增殖示踪荧光探针 25mg
MX3010-500T CFDA SE Cell Proliferation and Cell Tracking Kit 500T
MX3011-50UG Calcein, AM, Ultra Pure Grade钙黄绿素(绿色) 50μg
MX3012-500T Calcein AM/PI Double Stain Kit活细胞/死细胞双染试剂盒 500T
MX4101-1G Fluorescein diacetate (FDA)二乙酸荧光素 1g
MX4102-25MG CDCFDA, SE活细胞示踪探针 25mg
MX4103-100MG CDCFDA (5(6)-Carboxy-2’,7’-dichlorofluorescein diacetate)活细胞示踪探针 100mg
MX4104-25MG CFDA (5(6)-Carboxyfluorescein diacetate)活细胞示踪探针 25mg

CDCFDA (5(6)-Carboxy-2’,7’-dichlorofluorescein diacetate) 活细胞示踪探针


描述

CDCFDA (5(6)-Carboxy-2’,7’-dichlorofluorescein diacetate)

 

产品关键词:

CDCFDA;5(6)-CFDA;Fluorescein diacetate (FDA) 二乙酸荧光素;;Calcein AM钙黄绿素; Bacterial activity 细菌活性检测; CAS NO:127770-45-0;

产品信息

产品名称

产品编号 规格 价格(元)
CDCFDA (5(6)-Carboxy-2’,7’-dichlorofluorescein diacetate) 活细胞示踪探针 MX4103-100MG 100mg

1552

产品描述

CDCFDA(5(6)-Carboxy-2’,7’-dichlorofluorescein diacetate,CAS:127770-45-0),是一种胺反应的二乙酸荧光素(FDA)衍生物,能用来制备各种FDA偶联物。

CDCFDA本身无荧光,一旦扩散进入细胞后,被胞内非特异的酯酶水解生成一种荧光探针。这一探针专一聚集在具完整细胞膜结构的细胞内。CDCFDA不会对死细胞染色。CDCFDA标记可通过流式或荧光显微镜来检测。CDCFDA也常用来检测活细胞内的活性氧产生。

产品特性

1) CAS NO:127770-45-0

2) 同义名:5(6)- CDCFDA; 5(6)- CDCFDA, mixed isomers; 5(6)-Carboxy-2’,7’-dichlorofluorescein diacetate, mixed isomers; 5(6)-Carboxy-DCFDA;

3) 分子式:C25H14Cl2O9

4) 分子量:529.28 g/mol

5) 外观:白色至类白色固体

6) EX/Em:504/529nm

7) 溶解性:溶于DMSO

8) 化学结构式:

保存与运输方法

保存:-20℃避光干燥保存,至少1年有效。

运输:室温运输。

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。

相关产品

货号 名称 规格
MX3009-25MG CFDA, SE (CFSE) 细胞增殖示踪荧光探针 25mg
MX3010-500T CFDA SE Cell Proliferation and Cell Tracking Kit 500T
MX3011-50UG Calcein, AM, Ultra Pure Grade 钙黄绿素(绿色) 50μg
MX3012-500T Calcein AM/PI Double Stain Kit 活细胞/死细胞双染试剂盒 500T
MX4101-1G Fluorescein diacetate (FDA) 二乙酸荧光素 1g
MX4102-25MG CDCFDA, SE 活细胞示踪探针 25mg
MX4103-100MG CDCFDA (5(6)-Carboxy-2’,7’-dichlorofluorescein diacetate) 活细胞示踪探针 100mg
MX4104-25MG CFDA (5(6)-Carboxyfluorescein diacetate) 活细胞示踪探针 25mg

 

 

Fluorescein diacetate (FDA) 二乙酸荧光素


描述

Fluorescein diacetate (FDA) 二乙酸荧光素

产品关键词:

Fluorescein diacetate (FDA) 二乙酸荧光素;CFDA, SE (CFSE);Calcein AM钙黄绿素;CDCFDA; PKH26;CAS NO:596-09-8;

产品信息

产品名称 产品编号 规格 价格(元)
Fluorescein diacetate (FDA) 二乙酸荧光素 MX4101-1G 1g 150
Fluorescein diacetate (FDA) 二乙酸荧光素 MX4101-5G 5g 420

产品描述

二乙酸荧光素(Fluorescein diacetate, 简称FDA,CAS:596-09-8),是一种非荧光的疏水荧光素衍生物,能主动扩散进入细胞膜(包括哺乳动物细胞和细菌细胞),一旦进入细胞,经胞内酯酶水解二乙酸基团生成强荧光产物荧光素(Ex/Em=495/520nm)。此荧光素累积在含完整细胞膜的细胞内,从而用作细胞活力的指示标志物。而不具有完整细胞膜或缺乏活力代谢的细胞都无法聚集此荧光素,从而不会呈绿色荧光。FDA可能与碘化丙啶(PI)共染色,因死细胞能摄取PI,细胞被染成红色;而活细胞不能摄取PI,只能被染成绿色。这一双色分离死细胞和活细胞的方法比单色染色提供了一种相对更精确的活细胞定量分析。

产品特性

1) CAS NO:596-09-8

2) 化学名:Spiro(isobenzofuran-1(3H),9′-(9H)-xanthen)-3-one, 3′,6′-bis(acetyloxy)

3) 同义名:Cellstain-FDA; Di-O-acetylfluorescein; 3,6-Diacetoxyfluoran;荧光素二乙酸酯;荧光素双醋酸酯;

4) 分子式:C24H16O7

5) 分子量:416.39 g/mol

6) 外观:白色至淡黄色至深黄色粉末

7) 纯度:≥97%

8) 溶解性:溶于氯仿,乙醇,DMSO,水

9) 化学结构式:

保存与运输方法

保存:-20℃避光干燥保存,2年有效。

运输:室温运输。

使用方法

A.储存液配制

取低温保存的FDA粉末,置于室温回温至少20min。称取适量FDA(Mw:416.39 g/mol)用高质量无水DMSO充分溶解,配制成10mM 储存液。比如50mg FDA用12ml DMSO充分溶解混匀,即得到10mM 储存液。按照单次用量分装置于≤-20℃避光保存,避免反复冻融,至少2个月有效。

B.工作液配制

实验前,用HHBS缓冲液或其他生理缓冲液,pH 7.0稀释DMSO储存液到1-20µM 染色工作液,漩涡混匀。

C.染色步骤

1)用待测化合物处理细胞一段时间;

2)离心收集并调整得到2-10×105细胞/管;

3)用500µl染色工作液重悬细胞;

4)室温或37℃避光孵育15-30min;

5)吸掉染色工作液,用HHBS或其他缓冲液清洗细胞。用500µl预热的HHBS或培养基重悬细胞使其密度为2-10×105细胞/管;

6)用流式细胞仪(FL1通道)或荧光显微镜监测荧光变化(Ex/Em=490/520nm)。

注意事项

1) 本品保存过程中远离氧化剂。

2) 为了您的安全和健康,请穿实验服并戴一次性手套操作。

相关产品

货号 名称 规格
MX3009-25MG CFDA, SE (CFSE)细胞增殖示踪荧光探针 25mg
MX3010-500T CFDA SE Cell Proliferation and Cell Tracking Kit 500T
MX3011-50UG Calcein, AM, Ultra Pure Grade钙黄绿素(绿色) 50μg
MX3012-500T Calcein AM/PI Double Stain Kit活细胞/死细胞双染试剂盒 500T
MX4101-1G Fluorescein diacetate (FDA)二乙酸荧光素 1g
MX4102-25MG CDCFDA, SE活细胞示踪探针 25mg
MX4103-100MG CDCFDA (5(6)-Carboxy-2’,7’-dichlorofluorescein diacetate)活细胞示踪探针 100mg
MX4104-25MG CFDA (5(6)-Carboxyfluorescein diacetate)活细胞示踪探针 25mg

Fluorescein diacetate (FDA) 二乙酸荧光素


描述

Fluorescein diacetate (FDA) 二乙酸荧光素

产品关键词:

Fluorescein diacetate (FDA) 二乙酸荧光素;CFDA, SE (CFSE);Calcein AM钙黄绿素;CDCFDA; PKH26;CAS NO:596-09-8;

产品信息

产品名称 产品编号 规格 价格(元)
Fluorescein diacetate (FDA) 二乙酸荧光素 MX4101-1G 1g 150
Fluorescein diacetate (FDA) 二乙酸荧光素 MX4101-5G 5g 420

产品描述

二乙酸荧光素(Fluorescein diacetate, 简称FDA,CAS:596-09-8),是一种非荧光的疏水荧光素衍生物,能主动扩散进入细胞膜(包括哺乳动物细胞和细菌细胞),一旦进入细胞,经胞内酯酶水解二乙酸基团生成强荧光产物荧光素(Ex/Em=495/520nm)。此荧光素累积在含完整细胞膜的细胞内,从而用作细胞活力的指示标志物。而不具有完整细胞膜或缺乏活力代谢的细胞都无法聚集此荧光素,从而不会呈绿色荧光。FDA可能与碘化丙啶(PI)共染色,因死细胞能摄取PI,细胞被染成红色;而活细胞不能摄取PI,只能被染成绿色。这一双色分离死细胞和活细胞的方法比单色染色提供了一种相对更精确的活细胞定量分析。

产品特性

1) CAS NO:596-09-8

2) 化学名:Spiro(isobenzofuran-1(3H),9′-(9H)-xanthen)-3-one, 3′,6′-bis(acetyloxy)

3) 同义名:Cellstain-FDA; Di-O-acetylfluorescein; 3,6-Diacetoxyfluoran;荧光素二乙酸酯;荧光素双醋酸酯;

4) 分子式:C24H16O7

5) 分子量:416.39 g/mol

6) 外观:白色至淡黄色至深黄色粉末

7) 纯度:≥97%

8) 溶解性:溶于氯仿,乙醇,DMSO,水

9) 化学结构式:

保存与运输方法

保存:-20℃避光干燥保存,2年有效。

运输:室温运输。

使用方法

A.储存液配制

取低温保存的FDA粉末,置于室温回温至少20min。称取适量FDA(Mw:416.39 g/mol)用高质量无水DMSO充分溶解,配制成10mM 储存液。比如50mg FDA用12ml DMSO充分溶解混匀,即得到10mM 储存液。按照单次用量分装置于≤-20℃避光保存,避免反复冻融,至少2个月有效。

B.工作液配制

实验前,用HHBS缓冲液或其他生理缓冲液,pH 7.0稀释DMSO储存液到1-20µM 染色工作液,漩涡混匀。

C.染色步骤

1)用待测化合物处理细胞一段时间;

2)离心收集并调整得到2-10×105细胞/管;

3)用500µl染色工作液重悬细胞;

4)室温或37℃避光孵育15-30min;

5)吸掉染色工作液,用HHBS或其他缓冲液清洗细胞。用500µl预热的HHBS或培养基重悬细胞使其密度为2-10×105细胞/管;

6)用流式细胞仪(FL1通道)或荧光显微镜监测荧光变化(Ex/Em=490/520nm)。

注意事项

1) 本品保存过程中远离氧化剂。

2) 为了您的安全和健康,请穿实验服并戴一次性手套操作。

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FDA [Fluorescein diacetate],596-09-8

荧光素二乙酸酯(FDA)是一种用于活体染色的荧光探针,它是由人类Pi-类谷胱甘肽S-转移酶(hGSTP1)的酯水解活性在各种细胞溶质GST中选择性地激活的。

产品名称 FDA [Fluorescein diacetate],596-09-8
目录号 910PEG
中文名称 二乙酸荧光素;荧光素二乙酸酯
英文名称 Fluorescein diacetate
CAS 596-09-8
分子式 C24H16O7
分子量 416.3
存储条件 -20°干燥,避光
保存时间 一年
Ex/Em(nm) 488/530