Di-4-ANEPPS |
Catalog No.GC43435
One-Click Copy Product Info
|
Di-4-ANEPPS est un colorant sensible À la tension dans le potentiel de membrane.
Products are for research use only. Not for human use. We do not sell to patients.
Cas No.: 90134-00-2
Sample solution is provided at 25 µL, 10mM.
Di-4-ANEPPS is a fast-response membrane potential dye used to detect changes in membrane potential[1]. Di-4-ANEPPS fluoresces to potential changes through very rapid structural changes, so they are suitable for detecting fast and transient potential changes in cells. Di-4-ANEPPS is rapidly internalized in cells and is therefore mainly used in short-term experiments[2]. Di-4-ANEPPS is a highly water-soluble microinjection, the fluorescence is not affected by cell and tissue types, and can be used to measure membrane potential in various tissues and cell types as well as in artificial membranes. Di-4-ANEPPS is a common tool for mapping cardiac electrical activity[3].
References:
[1]. Maria Pura Hortigon-Vinagre, et al. The Use of Voltage Sensitive Dye di-4-ANEPPS and Video-Based Contractility Measurements to Assess Drug Effects on Excitation-Contraction Coupling in Human-Induced Pluripotent Stem Cell-Derived Cardiomyocytes. 2021 Mar 1;77(3):280-290. doi: 10.1097/FJC.0000000000000937.
[2]. LM Loew - Pure and Applied Chemistry. Potentiometric dyes: imaging electrical activity of cell membranes.1996 - degruyter.com
[3]. Marina Ronzhina, et al. Di-4-ANEPPS Modulates Electrical Activity and Progress of Myocardial Ischemia in Rabbit Isolated Heart. 2021 Jun 10;12:667065. doi: 10.3389/fphys.2021.667065. eCollection 2021.
This protocol provides a guide only and should be modified according to your specific needs.
1. Preparation of cell membrane staining solution
(1) Prepare DMSO or EtOH storage solution: the storage solution is prepared with DMSO or EtOH, the concentration is 1~5mM.
Note: Store unused stock solutions in aliquots at -20°C, avoid repeated freezing and thawing.
(2) Preparation of working solution: Dilute the stock solution with a suitable buffer (such as: serum-free medium, HBSS or PBS) to prepare a working solution with a concentration of 0.5-10μM.
Note: The final concentration of the working solution is prepared according to the experiences of different cells and experiments. Optimum conditions can be found from more than ten times the recommended concentration.
2. Suspension Cell Staining
(1) The suspended cells were centrifuged at 4°C, 1000-1500rpm for 3-5 minutes, and the supernatant was discarded. Wash twice with PBS for 5 minutes each.
(2) Add 1 mL of Di-4-ANEPPS working solution and incubate at room temperature for 5-30 minutes. The optimal incubation time is different for different cells.
(3) After the incubation, centrifuge at 1000-1500rpm for 5 minutes, remove the supernatant, add PBS to wash 2-3 times, 5 minutes each time.
(4) Resuspend the cells with pre-warmed serum-free cell culture medium or PBS. Observed by fluorescence microscopy or flow cytometry.
3. Staining of Adhesive Cells
(1) Culture adherent cells on sterile coverslips.
(2) Remove the coverslip from the culture medium, suck out the excess medium, and place the coverslip in a humid environment.
(3) Add 100uL of dye working solution from one corner of the cover slip and shake gently to make the dye evenly cover all the cells.
(4) Incubate at room temperature for 5-30 minutes, and the optimal incubation time is different for different cells.
(5) Discard the dye working solution, and wash the coverslip 2-3 times with culture medium.
4. Microscopic examination: the excitation/emission light of Di-4-ANEPPS are 495/705nm respectively.
Note:
1) Fluorescent dyes have quenching problems, please try to avoid light to slow downfluorescence quenching.
2) For your safety and health, please wear a lab coat and disposable gloves for operation.
| Cas No. | 90134-00-2 | SDF | |
| Canonical SMILES | CCCCN(CCCC)C(C=C1)=CC2=C1C=C(/C=C/C3=CC=[N+](CCCS([O-])(=O)=O)C=C3)C=C2 | ||
| Formula | C28H36N2O3S | M.Wt | 480.7 |
| Solubility | Soluble in DMSO | 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. |
||
| Shipping Condition | Evaluation sample solution: shipped with blue ice. All other sizes available: with RT, or with Blue Ice upon request. | ||
| Prepare stock solution | |||
|
1 mg | 5 mg | 10 mg |
| 1 mM | 2.0803 mL | 10.4015 mL | 20.803 mL |
| 5 mM | 416.1 μL | 2.0803 mL | 4.1606 mL |
| 10 mM | 208 μL | 1.0401 mL | 2.0803 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: >98.00% 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.)