Green CMFDA (Synonyms: 5-Chloromethylfluorescein diacetate) |
Catalog No.GC43783
One-Click Copy Product Info
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Green CMFDA is a cell-permeable fluorescent probe (Ex=485nm; Em=514nm) and can be used as a cell tracer.
Products are for research use only. Not for human use. We do not sell to patients.
Cas No.: 136832-63-8
Sample solution is provided at 25 µL, 10mM.
Green CMFDA is a cell-permeable fluorescent probe (Ex=485nm; Em=514nm) and can be used as a cell tracer[1]. Upon entering a living cell, Green CMFDA is cleaved by non-specific esterases common to living cells, producing a fluorescent compound, fluorescein, which accurately reflects changes associated with dormancy, and can be useful in laboratory experiments utilizing symbiont-bearing foraminifers[2]. Green CMFDA staining reduces the viability of human bone marrow-derived mesenchymal stem cells (hBM-MSCs) and causes changes in cell morphology, accompanied by a decrease in metabolic activity[3]. The Green CMFDA has been widely used for labeling living co-cultures, facilitating real-time imaging analysis and phagocytosis measurement[4].
References:
[1] Markovic J, Borrás C, Ortega A, et al. Glutathione is recruited into the nucleus in early phases of cell proliferation[J]. Journal of Biological Chemistry, 2007, 282(28): 20416-20424.
[2] Ross B J, Hallock P. Challenges in using CellTracker Green on foraminifers that host algal endosymbionts[J]. PeerJ, 2018, 6: e5304.
[3] Andrzejewska A, Jablonska A, Seta M, et al. Labeling of human mesenchymal stem cells with different classes of vital stains: robustness and toxicity[J]. Stem Cell Research & Therapy, 2019, 10(1): 187.
[4] Rodriguez-Manzanet R, Sanjuan M A, Wu H Y, et al. T and B cell hyperactivity and autoimmunity associated with niche-specific defects in apoptotic body clearance in TIM-4-deficient mice[J]. Proceedings of the National Academy of Sciences, 2010, 107(19): 8706-8711.
Cell labelling using Green CMFDA
This protocol only provides a guideline and should be modified according to your specific needs
1. Stock and dilute solution preparation for staining of cells
(1) Prepare Green CMFDA stock solution (10mM) by dissolving Green CMFDA powder in DMSO, which was then aliquoted into appropriate volumes and stored at -20°C.
(2) Dilute the Green CMFDA stock solution with PBS to a final 1µM concentration.
NOTE: The working solution must be prepared fresh on the day of the experiment.
2. Cell Preparation
Seed cultured cells in a 6-well plate or a 60-mm culture dish at 37°C, depending on the scale of the experiment.
3. Labeling of cells and analysis
(1) Wash the cells twice with PBS.
(2) Add Green CMFDA in PBS at a final concentration of 1μM, and incubate at 37°C for 30 minutes in the dark with gentle agitation.
(3) Collect cells into a 1.5ml tube and centrifuge at 500g for 10 minutes and discard the supernatant.
(4) Resuspend cells in culture medium and analyze using flow cytometry or fluorescence microscopy.
4. Experimental Precautions
(1) Ensure that the Green CMFDA is adequately protected from light.
(2) This protocol and all reagents are for scientific research use only.
References:
[1] Rodriguez-Manzanet R, Sanjuan M A, Wu H Y, et al. T and B cell hyperactivity and autoimmunity associated with niche-specific defects in apoptotic body clearance in TIM-4-deficient mice[J]. Proceedings of the National Academy of Sciences, 2010, 107(19): 8706-8711.
| Cas No. | 136832-63-8 | SDF | |
| Synonyms | 5-Chloromethylfluorescein diacetate | ||
| Chemical Name | 3',6'-bis(acetyloxy)-5-(chloromethyl)-spiro[isobenzofuran-1(3H),9'-[9H]xanthen]-3-one | ||
| Canonical SMILES | O=C(O1)C2=C(C=CC(CCl)=C2)C31C4=CC=C(OC(C)=O)C=C4OC5=C3C=CC(OC(C)=O)=C5 | ||
| Formula | C25H17ClO7 | M.Wt | 464.9 |
| Solubility | 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 | Storage | Store at -20°C, protect from light |
| General tips | Please select the appropriate solvent to prepare the stock solution according to the
solubility of the product in different solvents; once the solution is prepared, please store it in
separate packages to avoid product failure caused by repeated freezing and thawing.Storage method
and period of the stock solution: When stored at -80°C, please use it within 6 months; when stored
at -20°C, please use it within 1 month. To increase solubility, heat the tube to 37°C and then oscillate in an ultrasonic bath for some time. |
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| Shipping Condition | Evaluation sample solution: shipped with blue ice. All other sizes available: with RT, or with Blue Ice upon request. | ||
| Prepare stock solution | |||
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1 mg | 5 mg | 10 mg |
| 1 mM | 2.151 mL | 10.755 mL | 21.51 mL |
| 5 mM | 430.2 μL | 2.151 mL | 4.302 mL |
| 10 mM | 215.1 μL | 1.0755 mL | 2.151 mL |
Step 1: Enter information below (Recommended: An additional animal making an allowance for loss during the experiment)
g
μL
Step 2: Enter the in vivo formulation (This is only the calculator, not formulation. Please contact us first if there is no in vivo formulation at the solubility Section.)
Calculation results:
Working concentration: mg/ml;
Method for preparing DMSO master liquid: mg drug pre-dissolved in μL DMSO ( Master liquid concentration mg/mL, Please contact us first if the concentration exceeds the DMSO solubility of the batch of drug. )
Method for preparing in vivo formulation: Take μL DMSO master liquid, next addμL PEG300, mix and clarify, next addμL Tween 80, mix and clarify, next add μL saline, mix and clarify.
Method for preparing in vivo formulation: Take μL DMSO master liquid, next add μL Corn oil, mix and clarify.
Note: 1. Please make sure the liquid is clear before adding the next solvent.
2. Be sure to add the solvent(s) in order. You must ensure that the solution obtained, in the previous addition, is a clear solution before proceeding to add the next solvent. Physical methods such as vortex, ultrasound or hot water bath can be used to aid dissolving.
3. All of the above co-solvents are available for purchase on the GlpBio website.
Quality Control & SDS
- View current batch:
- Purity: >99.50% Appearance: A solid
- COA (Certificate of Analysis)
- SDS (Safety Data Sheet)
- Datasheet
Average Rating: 5 (Based on Reviews and 10 reference(s) in Google Scholar.)