Green CMFDA

A cell-permeable fluorescent probe

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货号: ajci71808
CAS: 136832-63-8
别名: 5-氯甲基荧光素二乙酸酯,5-Chloromethylfluorescein diacetate
分子式: C25H17ClO7
分子量: 464.9
纯度: >98%
溶解度: 0.1 M Na2CO3: 5 mg/mL,DMF: 25 mg/mL,DMSO: 33 mg/mL,Ethanol: 25 mg/mL,PBS (pH 7.2): 20 μ g/ml
储存: Store at -20°C, protect from light
库存: 现货

Background

Green CMFDA, as a non-terminal, non-fluorescent probe, can be cleaved by non-specific esterases common to living cells, producing a fluorescent compound, fluorescein, visible using a fluorescent microscope^[1]. CMFDA is excited with a monochromator at 492 nm and is emissed at 548 nm^[2].

In vitro, there was a a positive chondrogenic process of hBM-MSC labeled with 10 μM CellTracker? Green CMFDA, indicating these cells could differentiate into chondrogenic cells similarly to controlled non-labeled cells^[3]. In vitro experiment it shown that 4 weeks after grafting stem cells, the green fluorescence of the originally CMFDA-labeled (10 μM) bladder epithelia was still detectable in all of the resulting glandular prostate epithelium^[4].

参考文献:

[1] Bernhard, et al. Comparison of two methods to identify live benthic foraminifera: a test between Rose Bengal and CellTracker Green with implications for stable isotope paleoreconstructions. Paleoceanography. 2006;21.

[2] Trontelj K, et al. Cell electrofusion visualized with fluorescence microscopy. J Vis Exp. 2010 Jul 1;(41):1991.

[3] Andrzejewska A, et al. Labeling of human mesenchymal stem cells with different classes of vital stains: robustness and toxicity. Stem Cell Res Ther. 2019 Jun 25;10(1):187.

[4] Li X, et al. Urothelial transdifferentiation to prostate epithelia is mediated by paracrine TGF-beta signaling. Differentiation. 2009 Jan;77(1):95-102.

绿色 CMFDA 作为一种非末端、非荧光探针，可以被活细胞常见的非特异性酯酶切割，产生荧光化合物荧光素，使用荧光显微镜可见^{[1]。 CMFDA 在 492 nm 处用单色器激发，并在 548 nm 处发射^[2]。}

在体外，用 10 μM CellTracker? Green CMFDA 标记的 hBM-MSC 有一个阳性软骨形成过程，表明这些细胞可以像对照的未标记细胞一样分化成软骨形成细胞^[3] .体外实验表明，移植干细胞 4 周后，在所有生成的腺体前列腺上皮细胞中仍可检测到最初 CMFDA 标记的 (10 μM) 膀胱上皮细胞的绿色荧光^[4]。

Protocol

Cell experiment [1]:
Cell lines	platelets
Preparation Method	Samples of resting platelets and platelets activated by 100 μM ADP were adjusted to the platelet concentration of 3.5~3.8 × 108/L with autologous plasma, and incubated for 30 minutes at 37°C in 10 μM CMFDA. After incubation, the cytosolic esterase-induced fluorescence of platelets was measured by laser confocal microscopy and also photograph was taken.
Reaction Conditions	for 30 minutes at 37°C in 10 μM
Applications	The observation with laser confocal microscopy on CMFDA-stained platelets shows that nearly all the resting platelets release bright green fluorescence, whereas the ADP activated platelets release no or little fluorescence.
Animal experiment [2]:
Animal models	NOD/SCID or NOD/SCID/β2Mnull mice
Preparation Method	To determine the optimal time of BMDC injection in relation to bleo injury, 5 × 106 CMFDA-labeled BMDC were washed, resuspended in Hanks' buffer with heparin (50 U/ml), and administered via tail vein injection to NOD/SCID or NOD/SCID/β2Mnull mice 1, 2, 3, or 4 days post-bleo.
Dosage form	5 × 10⁶ ;1, 2, 3, or 4 days
Applications	There was a 10- to 100-fold increase in the percentage of CMFDA+ cells seen in NOD/SCID/β2Mnull mice compared with NOD/SCID mice, with ～0.04% positive cells seen when cells were infused at day 4 and mice were killed at day 7 after bleo.
参考文献: [1] Wang J, et al. Correlation between the In Vitro Functionality of Stored Platelets and the Cytosolic Esterase-Induced Fluorescence Intensity with CMFDA. PLoS One. 2015 Sep 21;10(9):e0138509. [2] Liebler JM, et al. Retention of human bone marrow-derived cells in murine lungs following bleomycin-induced lung injury. Am J Physiol Lung Cell Mol Physiol. 2008 Aug;295(2):L285-92.