Calcein AM (Calcein acetoxymethyl ester)
mRNA synthesis
In vitro transcription of capped mRNA with modified nucleotides and Poly(A) tail
Tyramide Signal Amplification (TSA)
TSA (Tyramide Signal Amplification), used for signal amplification of ISH, IHC and IC etc.
Phos Binding Reagent Acrylamide
Separation of phosphorylated and non-phosphorylated proteins without phospho-specific antibody
Cell Counting Kit-8 (CCK-8)
A convenient and sensitive way for cell proliferation assay and cytotoxicity assay
SYBR Safe DNA Gel Stain
Safe and sensitive stain for visualization of DNA or RNA in agarose or acrylamide gels.
Inhibitor Cocktails
Protect the integrity of proteins from multiple proteases and phosphatases for different applications.
Calcein AM (Calcein acetoxymethyl ester),中文名称钙黄绿素乙酰氧基甲酯,能够轻易穿透活细胞膜,是一种对活细胞进行荧光标记的绿色荧光探针。Calcein AM是在Calcein (钙黄绿素)的基础上引入了乙酰氧基甲酯(AM)基团,增加了疏水性,因此能够轻易穿透活细胞膜。Calcein AM本身并无荧光,一旦进入细胞后被细胞内的酯酶水解生成具有强负电荷的不能通透细胞膜的极性分子钙黄绿素(Calcein),从而被滞留在细胞内,并发出强绿色荧光[1]。由于Calcein AM的细胞毒性非常低,几乎不会影响正常细胞的功能,而且对pH值敏感性低[2],因此是目前染活细胞的最理想荧光探针之一。
由于死细胞缺乏酯酶,Calcein AM仅用于对活细胞的活力测试和短期标记,而染核酸的红色荧光染料碘化丙啶(Propidium Iodide, PI)由于不能穿透活细胞的细胞膜而只能染细胞膜完整性被破坏的死细胞,Calcein和PI-DNA都可被490 nm激发,因此Calcein AM常常与碘化丙啶联合使用,同时对活细胞和死细胞进行荧光染色[3]。用545 nm激发,仅可观察到死细胞。
Calcein AM可以应用于大多数的哺乳动物细胞。也有研究显示Calcein AM可以用于某些植物细胞如拟南芥(Arabidopsis)的根边缘样细胞[4]及某些酵母如Pichia anomala和Saccharomyces cerevisiae[5]。某些寄生虫如Leishmania由于细胞膜组分的原因,Calcein AM不能进入活细胞,但却可以进入凋亡早期的寄生虫细胞,从而与PI联用用于检测凋亡早期的寄生虫[6]。由于真菌和细菌有细胞壁,会阻碍Calcein AM进入细胞,因此Calcein AM不适合用于真菌和细菌。
References:
[1] Giordano G, Hong S, Faustman EM, et al. Measurements of cell death in neuronal and glial cells. Methods Mol Biol. 2011;758:171-8.
[2] Neri S, Mariani E, Meneghetti A, et al. Calcein-acetyoxymethyl cytotoxicity assay: standardization of a method allowing additional analyses on recovered effector cells and supernatants. Clin Diagn Lab Immunol. 2001;8(6):1131–1135.
[3] Kuksin D, Kuksin CA, Qiu J, et al. Cellometer image cytometry as a complementary tool to flow cytometry for verifying gated cell populations. Anal Biochem. 2016 Jun 15;503:1-7.
[4] Vicré M, Santaella C, Blanchet S, et al. Root border-like cells of Arabidopsis. Microscopical characterization and role in the interaction with rhizobacteria. Plant Physiol. 2005;138(2):998–1008.
[5] Prudêncio C, Sansonetty F, Sousa MJ, et al. Rapid detection of efflux pumps and their relation with drug resistance in yeast cells. Cytometry. 2000 Jan 1;39(1):26-35.
[6] Basmaciyan L, Azas N, Casanova M. Calcein+/PI- as an early apoptotic feature in Leishmania. PLoS One. 2017 Nov 7;12(11):e0187756.
Physical Appearance | A crystalline solid |
Storage | Store at -20°C |
M.Wt | 994.86 |
Cas No. | 148504-34-1 |
Formula | C46H46N2O23 |
Solubility | Soluble in DMSO |
Chemical Name | tetrakis(acetoxymethyl) 2,2',2'',2'''-(((3',6'-diacetoxy-3-oxo-3H-spiro[isobenzofuran-1,9'-xanthene]-4',5'-diyl)bis(methylene))bis(azanetriyl))tetraacetate |
SDF | Download SDF |
Canonical SMILES | O=C1O[C@@]2(C3=CC=CC=C31)C(C=C4)=C(C(CN(CC(OCOC(C)=O)=O)CC(OCOC(C)=O)=O)=C4OC(C)=O)OC5=C2C=CC(OC(C)=O)=C5CN(CC(OCOC(C)=O)=O)CC(OCOC(C)=O)=O |
运输条件 | 蓝冰运输或根据您的需求运输。 |
一般建议 | 不同厂家不同批次产品溶解度各有差异,仅做参考。若实验所需浓度过大至产品溶解极限,请添加助溶剂助溶或自行调整浓度。溶液形式一般不宜长期储存,请尽快用完。 |