标签归档：荧光

荧光染料Quin-2AM(Synonyms: Quin-2 acetoxymethyl ester)

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荧光染料Dye Reagents Quin-2AM;(Synonyms: Quin-2 acetoxymethyl ester)

Quin-2AM 是 Ca2+ 的荧光螯合剂，对钙有很高的亲和性。

Quin-2AM(Synonyms: Quin-2 acetoxymethyl ester)

Quin-2AM Chemical Structure

CAS No. : 83104-85-2

规格 价格 是否有货
1 mg ￥3890 询问价格 货期

* Please select Quantity before adding items.

生物活性

Quin-2AM is a fluorecent Ca2+ chelator, with high affinity for calcium.

体外研究
(In Vitro)

Quin-2AM (20 µM) or A23187 (20 µM) drastically activate mTF activity without detectable effect on mTF synthesis; both of which show that sustained functional upregulation during 24 h culture does not enhance Marcks expression. Quin-2AM (20 µM) changes the non-detectable Marcks level in 24 h cultures[1].

MCE has not independently confirmed the accuracy of these methods. They are for reference only.
分子量

829.76

Formula

C38H43N3O18
CAS 号

83104-85-2
运输条件

Room temperature in continental US; may vary elsewhere.
储存方式

Please store the product under the recommended conditions in the Certificate of Analysis.
参考文献

  • [1]. Chu AJ, et al. Possible role of Marcks in the cellular modulation of monocytic tissue factor-initiated hypercoagulation. Br J Haematol. 2002 Aug;118(2):569-76.

荧光染料(±)-ANAP

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荧光染料Dye Reagents (±)-ANAP;

(±)-ANAP 是一种非天然氨基酸荧光探针，为 prodan 的类似物，能够增加环境敏感性。

(±)-ANAP

(±)-ANAP Chemical Structure

CAS No. : 1185251-08-4

规格 是否有货
100 mg ; 询价 ;
250 mg ; 询价 ;
500 mg ; 询价 ;

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(plusmn;)-ANAP 的其他形式现货产品:

(plusmn;)-ANAP hydrochloride

生物活性

(±)-ANAP is the unnatural amino acid analog of prodan, acts as a fluorescent probes, and enhances environmental sensitivity.

体外研究
(In Vitro)

(±)-ANAP is the amino acid analog of prodan, acts as a fluorescent probes, and enhances environmental sensitivity with comparable or increased brightness[1]. (±)-ANAP is sensitive to polarity with changes in intensity and emission wavelength[2].

MCE has not independently confirmed the accuracy of these methods. They are for reference only.
分子量

272.30

Formula

C15H16N2O3
CAS 号

1185251-08-4
运输条件

Room temperature in continental US; may vary elsewhere.
储存方式

Please store the product under the recommended conditions in the Certificate of Analysis.
参考文献

  • [1]. Lee HS, et al. Genetic incorporation of a small, environmentally sensitive, fluorescent probe into proteins in Saccharomyces cerevisiae. J Am Chem Soc. 2009 Sep 16;131(36):12921-3.

    [2]. Xiang Z, et al. Enantiospecific synthesis of genetically encodable fluorescent unnatural amino acid L-3-(6-acetylnaphthalen-2-ylamino)-2-aminopropanoic acid. J Org Chem. 2011 Aug 5;76(15):6367-71.

荧光染料ADHP(Synonyms: 10-Acetyl-3,7-dihydroxyphenoxazine)

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荧光染料Dye Reagents ADHP;(Synonyms: 10-Acetyl-3,7-dihydroxyphenoxazine)

ADHP 是一种荧光过氧化物酶底物（λex=530 nm, λem=590 nm）。

ADHP(Synonyms: 10-Acetyl-3,7-dihydroxyphenoxazine)

ADHP Chemical Structure

CAS No. : 119171-73-2

规格 价格 是否有货
5 mg ￥2200 询问价格 货期

* Please select Quantity before adding items.

生物活性

ADHP is a fluorogenic peroxidase substrate (λex=530 nm, λem=590 nm).

体外研究
(In Vitro)

To obtain the parameters Km and kcat for Compound I, two independent methods are used. Initially, the oxidation of ADHP using the injector functionality built-in to the fluorescence plate reader is studied. The auto-injector dispenses the H2O2 to initiate the reaction, as a means of generating a set of progress curves. Analysis for MPO-mediated oxidation of ADHP gives a Km of 31±4 μM and the kcat of 186± 6 s−1.The kobs also increases over the experimental range of ADHP concentrations from 1 to 80 μM and for the converse experiment holding substrate constant over 3 to 45 nM MPO. The apparent second order rate constant obtain from the slope of kobs against ADHP concentration Kappon is 2.1±0.2 mM/s[1].

MCE has not independently confirmed the accuracy of these methods. They are for reference only.
分子量

257.24

Formula

C14H11NO4
CAS 号

119171-73-2
中文名称

10-乙酰基-3,7-二羟基吩嗪
运输条件

Room temperature in continental US; may vary elsewhere.
储存方式

Please store the product under the recommended conditions in the Certificate of Analysis.
参考文献

  • [1]. Jiansheng Huang, et al. Ordered Cleavage of Myeloperoxidase Ester Bonds Releases Active site Heme Leading to Inactivation of Myeloperoxidase by Benzoic Acid Hydrazide Analogs. Arch Biochem Biophys. 2014 Apr 15; 548: 74–85.
Kinase Assay
[1]

ADHP, 4-ABAH, 2-ABAH, 4-BAH, 4-FBAH, 4-NBAH, 4-TFMBAH, 3-DMABAH, NaN3 and isoniazid are dissolved in DMSO and subsequently diluted into assay buffer. The final concentration of DMSO in the reaction is less than 0.5 % (v/v), which does not affect fluorescence of the oxidized ADHP product 7-hydroxyl-3H-phenoxazin-3-one (resorufin). Reactions of ADHP (20 μM) are incubated with MPO (2.8 nm) in assay buffer and initiated by the addition of 1/10th volume H2O2 from a serial dilution basin. To determine the effect that the simplest benzoic acid hydrazide inhibitor or its analog 4-TFMBAH has on the heme catalytic ability of MPO, MPO (1.2 μM) is incubated for 10 min with different concentrations of BAH inhibitor (0, 0.025, 0.25, 2.5, 12.5 and 25 mM) with ADHP (40 μM) and timing of the reaction is measured following addition of H2O2 (20 μM) ADHP. All reactions are measured in assay buffer at room temperature. Samples of 20 μL are added to non-reducing sample loading buffers, and then loaded without prior heating and resolved by 4-15% gradient SDS-polyacrylamide gel electrophoresis[1].

MCE has not independently confirmed the accuracy of these methods. They are for reference only.
参考文献

  • [1]. Jiansheng Huang, et al. Ordered Cleavage of Myeloperoxidase Ester Bonds Releases Active site Heme Leading to Inactivation of Myeloperoxidase by Benzoic Acid Hydrazide Analogs. Arch Biochem Biophys. 2014 Apr 15; 548: 74–85.

5(6)-Carboxyfluorescein(Synonyms: 5(6)-羧基荧光素 5(6)-FAM 5-(and-6)-Carboxyfluorescein mixed isomers)

发表于由

5(6)-Carboxyfluorescein;(Synonyms: 5(6)-羧基荧光素; 5(6)-FAM; 5-(and-6)-Carboxyfluorescein mixed isomers) 纯度: 99.00%

5(6)-Carboxyfluorescein (5(6)-FAM) 是一种胺反应性 pH 敏感的绿色荧光标记物。5(6)-Carboxyfluorescein 可用于标记蛋白质、多肽和氨基酸。5(6)-Carboxyfluorescein 可用于体内实验肿瘤位置检测。

5(6)-Carboxyfluoresceinamp;;(Synonyms: 5(6)-羧基荧光素; 5(6)-FAM; 5-(and-6)-Carboxyfluorescein mixed isomers)

5(6)-Carboxyfluorescein Chemical Structure

CAS No. : 72088-94-9

规格 价格 是否有货 数量
10;mM;*;1 mL in DMSO ￥500 In-stock
500 mg ￥400 In-stock
1 g ￥500 In-stock
5 g ; 询价 ;
10 g ; 询价 ;

* Please select Quantity before adding items.

5(6)-Carboxyfluorescein 相关产品

bull;相关化合物库:

  • Bioactive Compound Library Plus
  • Anti-Cancer Compound Library

生物活性

5(6)-Carboxyfluorescein (5(6)-FAM) is an amine-reactive pH-sensitive green fluorescent probe. 5(6)-Carboxyfluorescein (5(6)-FAM) can be used to label proteins, peptides and nucleotides. 5(6)-Carboxyfluorescein can be used for the detection of tumour areas in vivo[1][2].

体外研究
(In Vitro)

5(6)-Carboxyfluorescein has two main characteristics: it has two wavelengths of maximum absorbance (465 and 490 nm) and its fluorescence emission (maximum, 515 nm) increases as a function of pH in the physiological pH range of 6-7.4[2].

MCE has not independently confirmed the accuracy of these methods. They are for reference only.
体内研究
(In Vivo)

5(6)-Carboxyfluorescein (5 mg/kg; i.p.) can be used for in vivo pH mapping of tumor tissue[2].

MCE has not independently confirmed the accuracy of these methods. They are for reference only.

Animal Model: CDF mice, bearing lymphoid leukemia P388[2]
Dosage: 5 mg/kg
Administration: Injected intraperitoneally
Result: Could be used for measurement and imaging of tumor tissue.

分子量

752.63

Formula

C42H24O14
CAS 号

72088-94-9
中文名称

5(6)-羧基荧光素
运输条件

Room temperature in continental US; may vary elsewhere.
储存方式

-20deg;C, protect from light

*In solvent : -80deg;C, 6 months; -20deg;C, 1 month (protect from light)
溶解性数据
In Vitro:;

DMSO : ≥ 41 mg/mL (54.48 mM)

* “≥” means soluble, but saturation unknown.

配制储备液
浓度 溶剂体积 质量 1 mg 5 mg 10 mg
1 mM 1.3287 mL 6.6434 mL 13.2867 mL
5 mM 0.2657 mL 1.3287 mL 2.6573 mL
10 mM 0.1329 mL 0.6643 mL 1.3287 mL

*

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, 6 months; -20°C, 1 month (protect from light)。-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。
参考文献

  • [1]. Maksim V Kvach, et al. 5(6)-carboxyfluorescein revisited: new protecting group, separation of isomers, and their spectral properties on oligonucleotides. Bioconjug Chem. Sep-Oct 2007;18(5):1691-6.

    [2]. Wen Ma, et al. A Cell Membrane-Targeting Self-Delivery Chimeric Peptide for Enhanced Photodynamic Therapy and In Situ Therapeutic Feedback. Adv Healthc Mater. 2020 Jan;9(1):e1901100.

2-Aminopurine(Synonyms: 2-氨基嘌呤)

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2-Aminopurine;(Synonyms: 2-氨基嘌呤) 纯度: 99.53%

2-Aminopurine 是鸟苷和腺苷的荧光类似物，是一种广泛使用的基于荧光衰变的 DNA 结构探针。当将 2-Aminopurine 插入寡核苷酸时，其荧光会通过与天然碱基堆积而被高度淬灭。2-Aminopurine 已被用于探测核酸的结构和动力学。

2-Aminopurineamp;;(Synonyms: 2-氨基嘌呤)

2-Aminopurine Chemical Structure

CAS No. : 452-06-2

规格 价格 是否有货 数量
100 mg ￥500 In-stock
200 mg ; 询价 ;
500 mg ; 询价 ;

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2-Aminopurine 相关产品

bull;相关化合物库:

  • Bioactive Compound Library Plus
  • Nucleotide Compound Library

生物活性

2-Aminopurine, a fluorescent analog of guanosine and adenosine, is a widely used fluorescence-decay-based probe of DNA structure. When 2-Aminopurine is inserted in anoligonucleotide, its fluorescence is highly quenched by stacking with the natural bases. 2-Aminopurine has been used to probe nucleic acid structure and dynamics[1][2].

体外研究
(In Vitro)

2-Aminopurine (2AP) is not valuable as afluorescent label because its fluorescence is highly quenched by stacking with the natural bases, when it is inserted in anoligonucleotide. However, it is this very susceptibility to interbase quenching that makes 2AP an exquisitely sensitivefluorescent probe of nucleic acid structure[1].
2-Aminopurine differs from adenine (6-aminopurine) only in the position of the exocyclic amine group, and yet its fluorescence intensity is one thousand times that of adenine[1].

MCE has not independently confirmed the accuracy of these methods. They are for reference only.
分子量

135.13

Formula

C5H5N5
CAS 号

452-06-2
中文名称

2-氨基嘌呤
运输条件

Room temperature in continental US; may vary elsewhere.
储存方式

4deg;C, protect from light

*In solvent : -80deg;C, 6 months; -20deg;C, 1 month (protect from light)
溶解性数据
In Vitro:;

DMSO : 5 mg/mL (37.00 mM; Need ultrasonic)

H2O : 1.35 mg/mL (9.99 mM; Need ultrasonic)

配制储备液
浓度 溶剂体积 质量 1 mg 5 mg 10 mg
1 mM 7.4003 mL 37.0014 mL 74.0028 mL
5 mM 1.4801 mL 7.4003 mL 14.8006 mL
10 mM 0.7400 mL 3.7001 mL 7.4003 mL

*

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, 6 months; -20°C, 1 month (protect from light)。-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。
参考文献

  • [1]. J M Jean, et al. 2-Aminopurine fluorescence quenching and lifetimes: role of base stacking. Proc Natl Acad Sci U S A. 2001 Jan 2;98(1):37-41.

    [2]. Dehong Tan, et al. Decreased glycation and structural protection properties of γ-glutamyl-S-allyl-cysteine peptide isolated from fresh garlic scales (Allium sativum L.). Nat Prod Res. 2015;29(23):2219-22.

TO-PRO 1

发表于由

TO-PRO 1;

TO-PRO 1 是一种 DNA 结合荧光染料，附着在 Feraheme (FH) 纳米颗粒 (NP) 的表面，以获得荧光染料功能化的 NP。TO-PRO 1 嵌入结合 DNA，用作坏死细胞的重要荧光染料。

TO-PRO 1amp;;

TO-PRO 1 Chemical Structure

CAS No. : 157199-59-2

规格 价格 是否有货
5 mM * 50 μL in DMSO ￥5500 询问价格 货期

* Please select Quantity before adding items.

生物活性

TO-PRO 1 is a DNA binding fluorochrome, that atached to the surface of the Feraheme (FH) nanoparticle (NP), to obtain a fluorochrome-functionalized NP. TO-PRO 1 binds DNA through intercalation, and acts as a vital fluorochrome for necrotic cells[1].

分子量

645.38

Formula

C24H29I2N3S
CAS 号

157199-59-2
运输条件

Room temperature in continental US; may vary elsewhere.
储存方式

Please store the product under the recommended conditions in the Certificate of Analysis.
参考文献

  • [1]. Hoonsung Cho, et al. Fluorochrome-functionalized nanoparticles for imaging DNA in biological systems. ACS Nano. 2013 Mar 26;7(3):2032-41.

5-IAF(Synonyms: 5-碘乙酰氨基荧光素 5-Iodoacetamidofluorescein)

发表于由

5-IAF;(Synonyms: 5-碘乙酰氨基荧光素; 5-Iodoacetamidofluorescein) 纯度: ge;95.0%

5-IAF 是荧光素的偶氮乙酰胺衍生物。

5-IAFamp;;(Synonyms: 5-碘乙酰氨基荧光素; 5-Iodoacetamidofluorescein)

5-IAF Chemical Structure

CAS No. : 63368-54-7

规格 价格 是否有货 数量
5 mg ￥770 In-stock
10 mg ; 询价 ;
50 mg ; 询价 ;

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5-IAF 相关产品

bull;相关化合物库:

  • Bioactive Compound Library Plus
  • Anti-Cancer Compound Library

生物活性

5-IAF is an idoacetamide derivate of fluoresceine.

体外研究
(In Vitro)

8 µM GSH sample solution is derivatized with 8, 80, 200, 400, 800, 1600, 2000, 2500 µM 5-IAF, respectively and analyzed with CE-LIF. The highest signal intensity is obtained with 400 and 800 µM 5-IAF[2].

MCE has not independently confirmed the accuracy of these methods. They are for reference only.
分子量

515.25

Formula

C22H14INO6
CAS 号

63368-54-7
中文名称

5-碘乙酰氨基荧光素
运输条件

Room temperature in continental US; may vary elsewhere.
储存方式
Powder -20deg;C 3 years
In solvent -80deg;C 6 months
-20deg;C 1 month
溶解性数据
In Vitro:;

DMSO : 50 mg/mL (97.04 mM; Need ultrasonic)

配制储备液
浓度 溶剂体积 质量 1 mg 5 mg 10 mg
1 mM 1.9408 mL 9.7040 mL 19.4081 mL
5 mM 0.3882 mL 1.9408 mL 3.8816 mL
10 mM 0.1941 mL 0.9704 mL 1.9408 mL

*

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, 6 months; -20°C, 1 month。-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。

In Vivo:

请根据您的实验动物和给药方式选择适当的溶解方案。以下溶解方案都请先按照 In Vitro 方式配制澄清的储备液，再依次添加助溶剂：

——为保证实验结果的可靠性，澄清的储备液可以根据储存条件，适当保存；体内实验的工作液，建议您现用现配，当天使用； 以下溶剂前显示的百
分比是指该溶剂在您配制终溶液中的体积占比；如在配制过程中出现沉淀、析出现象，可以通过加热和/或超声的方式助溶

  • 1.

    请依序添加每种溶剂：;10% DMSO ;; 40% PEG300 ;; 5% Tween-80 ;; 45% saline

    Solubility: 1.43 mg/mL (2.78 mM); Suspended solution; Need ultrasonic

    此方案可获得 1.43 mg/mL (2.78 mM) 的均匀悬浊液，悬浊液可用于口服和腹腔注射。

    以 1 mL 工作液为例，取 100 μL 14.3 mg/mL 的澄清 DMSO 储备液加到 400 μL PEG300 中，混合均匀；向上述体系中加入50 μL Tween-80，混合均匀；然后继续加入 450 μL生理盐水定容至 1 mL。

    将 0.9 g 氯化钠，完全溶解于 100 mL ddH₂O 中，得到澄清透明的生理盐水溶液

  • 2.

    请依序添加每种溶剂：;10% DMSO ;; 90% (20% SBE-β-CD in saline)

    Solubility: 1.43 mg/mL (2.78 mM); Suspended solution; Need ultrasonic

    此方案可获得 1.43 mg/mL (2.78 mM) 的均匀悬浊液，悬浊液可用于口服和腹腔注射。

    以 1 mL 工作液为例，取 100 μL 14.3 mg/mL 的澄清 DMSO 储备液加到 900 μL 20% 的 SBE-β-CD 生理盐水水溶液中，混合均匀。

    将 2 g 磺丁基醚 β-环糊精加入 5 mL 生理盐水中，再用生理盐水定容至 10 mL，完全溶解，澄清透明
*以上所有助溶剂都可在 MCE 网站选购。
参考文献

  • [1]. Salvatore Sotgia, et al. Plasma L-Ergothioneine Measurement by High-Performance Liquid Chromatography and Capillary Electrophoresis after a Pre-Column Derivatization with 5-Iodoacetamidofluorescein (5-IAF) and Fluorescence Detection. PLoS One. 2013; 8(7): e70374.

    [2]. Yan Wang, et al. Determination of free and protein-bound glutathione in HepG2 cells using capillary electrophoresis with laser-induced fluorescence detection. J Chromatogr A. 2009 Apr 17;1216(16):3533-7.
Kinase Assay
[2]

HepG2 cells are lysed in 1 mL 2% acetonitrile in water, and transfered to a 1.5 mL centrifuge tube. An additional 200 µL 2% acetonitrile is used to rinse the dish and combined with the cell extract. To measure the free GSH concentration, three aliquots of 80 µL cell extract are immediately transferred to 0.5 mL centrifuge tubes after a quick mix. Then 20 µL 50 µM NAC is added, followed by the addition of 100 µL 400 µM 5-IAF to the mixture. The derivatization is conducted at room temperature in dark for 1 h, after which the mixtures are centrifuged at 9000× g for 5 min to sediment the insoluble proteins. 10 µL supernatant is diluted 100× with water and analyzed immediately by CE-LIF[2].

MCE has not independently confirmed the accuracy of these methods. They are for reference only.
参考文献

  • [1]. Salvatore Sotgia, et al. Plasma L-Ergothioneine Measurement by High-Performance Liquid Chromatography and Capillary Electrophoresis after a Pre-Column Derivatization with 5-Iodoacetamidofluorescein (5-IAF) and Fluorescence Detection. PLoS One. 2013; 8(7): e70374.

    [2]. Yan Wang, et al. Determination of free and protein-bound glutathione in HepG2 cells using capillary electrophoresis with laser-induced fluorescence detection. J Chromatogr A. 2009 Apr 17;1216(16):3533-7.

5-Aminofluorescein(Synonyms: 5-氨基荧光素 5-AF)

发表于由

5-Aminofluorescein;(Synonyms: 5-氨基荧光素; 5-AF) 纯度: 99.76%

5-Aminofluorescein (5-AF) 是一种荧光标记物，共价结合到人血清白蛋。激发波长为 490 nm，发射波长为 515 nm。

5-Aminofluoresceinamp;;(Synonyms: 5-氨基荧光素; 5-AF)

5-Aminofluorescein Chemical Structure

CAS No. : 3326-34-9

规格 价格 是否有货 数量
Free Sample (0.1-0.5 mg) ; Apply now ;
10;mM;*;1 mL in DMSO ￥660 In-stock
500 mg ￥600 In-stock
1 g ￥880 In-stock
5 g ; 询价 ;
10 g ; 询价 ;

* Please select Quantity before adding items.

5-Aminofluorescein 相关产品

bull;相关化合物库:

  • Covalent Screening Library Plus
  • Bioactive Compound Library Plus
  • Covalent Screening Library

生物活性

5-Aminofluorescein (5-AF) is a fluorescence marker, covalently bound to human serum albumin. The excitation wavelength is 490 nm and the emission wavelength is 515 nm.

分子量

347.32

Formula

C20H13NO5
CAS 号

3326-34-9
中文名称

5-氨基荧光素
运输条件

Room temperature in continental US; may vary elsewhere.
储存方式

4deg;C, protect from light

*In solvent : -80deg;C, 6 months; -20deg;C, 1 month (protect from light)
溶解性数据
In Vitro:;

DMSO : ≥ 32 mg/mL (92.13 mM)

* “≥” means soluble, but saturation unknown.

配制储备液
浓度 溶剂体积 质量 1 mg 5 mg 10 mg
1 mM 2.8792 mL 14.3959 mL 28.7919 mL
5 mM 0.5758 mL 2.8792 mL 5.7584 mL
10 mM 0.2879 mL 1.4396 mL 2.8792 mL

*

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, 6 months; -20°C, 1 month (protect from light)。-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。
参考文献

  • [1]. Ding R, et al. Pharmacokinetics of 5-aminofluorescein-albumin, a novel fluorescence marker of brain tumors during surgery. J Clin Pharmacol. 2011 May;51(5):672-8.

    [2]. Miao R, et al. Facile method for modification of the silicon nanowires and its application in fabrication of pH-sensitive chips. ACS Appl Mater Interfaces. 2013 Mar 13;5(5):1741-6.

    [3]. Wang T, et al. Fluorescein Derivatives as Bifunctional Molecules for the Simultaneous Inhibiting and Labeling of FTO Protein. J Am Chem Soc. 2015 Nov 4;137(43):13736-9.

    [4]. Shan L, et al. Multi-small molecule conjugations as new targeted delivery carriers for tumor therapy. Int J Nanomedicine. 2015 Sep 1;10:5571-91.

Enzo使用荧光染料检测牛主动脉细胞的自噬情况

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Enzo使用荧光染料检测牛主动脉细胞的自噬情况

Enzo使用荧光染料检测牛主动脉细胞的自噬情况



细胞组分的降解对于维持体内平衡不可或缺，这个复杂的过程受到内溶酶体和自噬途径的调节。目前已经拥有多种试剂和报告系统研究这些途径，但是每种研究手段都有优点和局限性。最近Histochemistry and Cell Biology杂志上发表了一篇文章，Oeste和他的同事们使用Enzo的Cyto-ID® 和Lyso-ID® 染料产品检测牛主动脉内皮细胞(BAEC)中的自噬泡和内溶酶体泡。虽然他们的结果是在特定的细胞系和特定的实验条件下得到的，但是他们的发现有力证明了这些染料的使用优势，可替代或者与LC3报告系统中的染料(如monodansyl cadaverine, MDC)联用。其结果如下： 

Lyso-ID信号可与LampI-GFP 和GFP8共定位，这个GFP结构包含内溶酶体靶向识别RhoB GTP酶的信号。当用chloroquine或干扰内涵体稳态的U18666A处理细胞时，共定位会持续。

·         使用RFP-LC3转染的牛主动脉内皮细胞，利用氨基酸饥饿法诱导其细胞发生自噬后，研究人员展示了Cyto-ID自噬染料与RFP-LC3的共定位结果图。尤其是当加入溶酶体蛋白酶抑制剂抑制RFP-LC3的降解，这种共定位更为明显。饥饿处理过程中，Cyto-ID信号会早于RFP-LC3的点状信号之前形成，这为自噬泡形成提供了早期检测。在饥饿期间，Cyto-ID信号会对自噬诱导剂Rapamycin及抑制剂3-methyladenine (3-MA)产生应答。

·         作者指出相比于MDC带有显著本底而没有自噬诱导的信号，Cyto-ID减少了背景色。Bampton及其同事(2005)早先曾提出，MDC会因离子捕获而在酸性部位积累，导致背景信号显著。伴随着自噬的诱导， 只有部分MDC和Cyto-ID信号重叠，反映了Cyto-ID染料标记更为特异。

·         结合应用Cyto-ID和Lyso -ID染料，Oeste和同事实验结果中预测了来源于噬溶酶体融合的共同定位信号，以及Cyto-ID和Lyso -ID染色细胞器的独特亚群。

 

Enzo为自噬研究提供了多种产品，包括活细胞分析试剂盒、免疫检测、化合物库和抗体，以下是其中一些产品的简介。

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产品

特点

分析试剂盒

Lyso-ID® Red detection kit (GFP-Certified®)

适用于溶酶体和自生性溶酶体活细胞成像。长波红光易于与其它荧光探针联用，如荧光素，香豆素和GFP。极耐光漂白、不易猝灭。不会转成绿色荧光，不会发射别的荧光。

Lyso-ID® Green detection kit

该试剂盒是专门为表达RFP细胞株和罗丹明类荧光团设计的，蓝色、青色或橙黄色荧光蛋白（BFPs，CFPs，OFPs个）也可以使用。

Lyso-ID® Red cytotoxicity kit (GFP-Certified®) for microplates

快速，10-15分钟染料孵育、高通量分析。在短短的3小时内即可获得定量结果，包含药物处理过程。

Cyto-ID® Autophagy detection kit

简单、无需转染即可定量监测活细胞自噬。

PROTEOSTAT® Aggresome detection kit

定量检测错误折叠的蛋白，与神经退行性疾病、肝脏疾病和毒理学的研究相关。

p62 ELISA kit

用于自噬研究，高效、定量的p62 ELISA试剂盒，可检测低至100 pg/ml的p62进行。

NBR1 ELISA kit

率先投放市场，用于自噬研究的NBR1定量检测试剂盒，可检测65 pg/ml的 NBR1。

SCREEN-WELL® Autophagy library

不同结构和机制的化合物，用于胞内和体外研究，促进/抗自噬分子的作用机制。

抗体

LC3, mAb (2G6) (fluorescein labeled)

识别所有单体（LC3-I）和PE交联（LC3-II）形式的人LC3/GABARAP 蛋白 (Atg8 同系物) 。

LC3, mAb (5H3)

识别人LC3。识别两种形式的内源性LC3。

LC3B, mAb (2G6)

识别人、小鼠、大鼠、猴和仓鼠LC3B。识别两种形式的内源性LC3B。

LC3B, mAb (5F10)

识别人、小鼠、大鼠、狗和仓鼠LC3B。识别这两种形式的内源性LC3B。

小分子

3-Methyladenine

PI3激酶抑制剂，著名的自噬抑制剂。

Rapamycin

大环-三烯抗生素具有强效免疫抑制剂的活性。它与FKBP12形成的复合物结合一个效应子，从而抑制IL-2和其它生长因子。

U-18666A

可抑制胆固醇从内涵体/溶酶体运输至内质网的化合物。

References:

 

·　Oeste CL, et al. Interactions between autophagic and endo-lysosomal markers in endothelial cells. Histochem. Cell. Biol. (2012) (epub ahead of print).

·　Bampton ET, et al. The dynamics of autophagy visualized in live cells: from autophagosome formation to fusion with endo/lysosomes. Autophagy. (2005) 1:23–36.

·　Niemann A, et al. The lysosomotropic agent monodansylcadaverine also acts as a solvent polarity probe. J Histochem Cytochem (2000) 48:251-258.

·　Stenmark H (2009) Rab GTPases as coordinators of vesicle traffic. Nat Rev Mol Cell Biol (2009) 10:513-525.

Enzo使用荧光染料检测牛主动脉细胞的自噬情况

BES 特异性荧光探针

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BES 特异性荧光探针

  • 产品特性
  • 相关资料
  • Q&A
  • 参考文献

BES 特异性荧光探针BES 特异性荧光探针

超氧化物特异性荧光探针 BES-So-AM / BES-So

◆特点


●  比氢化乙啡啶（hydroethidine）有更高的特异性

●  与超氧化物特异性反应，用于监测活细胞内超氧化物水平

●  BES-So-AM 可穿透细胞膜，无需打孔

●  几乎不溶于水，可制成水溶液使用

●  可用于荧光显微镜、流式细胞仪


BES 特异性荧光探针


■ 细胞通透性


BES 特异性荧光探针

产品编号 产品名称 Ex/Em(nm)
021-17801 BES-So-AM 505/544





■ 无细胞通透性


产品编号 产品名称 Ex/Em(nm)
025-18921 BES-So 505/544