Diphenyl Blue (Direct Blue 14) |
| Catalog No.GC30152 |
Diphenyl Blue (Trypan blue), a cell viability dye, is the most commonly used dye to identify dead cells and is used to test cell membrane integrity and cell viability
Products are for research use only. Not for human use. We do not sell to patients.
Cas No.: 72-57-1
Sample solution is provided at 25 µL, 10mM.
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Diphenyl Blue (Trypan blue), a cell viability dye, is the most commonly used dye to identify dead cells and is used to test cell membrane integrity and cell viability[1]. Normal living cells have a complete membrane structure, which can repel trypan blue and prevent it from entering the cell; while cells with inactive or incomplete membranes have increased membrane permeability and can be stained blue by trypan blue It is generally considered that the integrity of the cell membrane is lost, that is, the cell is considered to be dead, which is opposite to the effect of neutral red. Therefore, with the help of trypan blue staining, it is very simple and fast to distinguish living cells from dead cells. Trypan blue is one of the most commonly used stains for dead cell identification in tissue and cell culture[1]. Macrophages are able to phagocytose diphenyl blue, so it can be used as a live stain for macrophages[2].
References:
[1]. B A Avelar-Freitas,et. Trypan blue exclusion assay by flow cytometry. 2014 Apr;47(4):307-15. doi: 10.1590/1414-431X20143437. Epub 2014 Mar 18.
[2]. Daly, M. L., DeRosa, C. A., Kerr, C., Morris, W. A., & Fraser, C. L. (2016). Blue thermally activated delayed fluorescence from a biphenyl difluoroboron β-diketonate. RSC Advances, 6(85), 81631–81635. doi:10.1039/c6ra18374c.
This protocol provides a guide only and should be modified according to your specific needs.
Ⅰ. Preparation of Diphenyl Blue (Trypan Blue) Dye Solution
1. Prepare diphenyl blue (trypan blue) stock solution: Dissolve 10 mg of trypan blue in 100 mL of 0.85% Nacl to obtain a 0.4% trypan blue stock solution.
Note: It is recommended to store the stock solution in aliquots at -20°C-80°C in the dark, and avoid repeated freeze-thaw cycles.
2. Prepare diphenyl blue (trypan blue) working solution: Use serum-free cell culture medium or PBS to dilute the stock solution to prepare trypan blue working solution with a concentration of 0.04%.
Note: Please adjust the concentration of the trypan blue working solution according to the experimental needs, and prepare it now.
Ⅱ. Cell staining
1. Suspended cells: Collect cells by centrifugation, add PBS to wash twice, 5 minutes each time.
Adherent cells: remove medium and add trypsin to digest cells. Discard the supernatant after centrifugation, add PBS to wash twice, 5 minutes each time.
2. Add 1mL trypan blue working solution and incubate for 5 minutes at room temperature.
3. Centrifuge at 400g for 3-4 minutes at 4°C, discard the supernatant, add PBS to wash the cells twice, 5 minutes each time.
4. Resuspend the cells with 1 mL of serum-free medium or PBS. Cell viability was calculated by counting under a microscope or after taking pictures under a microscope[3].
Cell survival rate (%)=total number of live cells/(total number of live cells+total number of dead cells)*100%
3. Matters needing attention:
1. The staining time should not be too long, otherwise the living cells will gradually accumulate the dye and become stained, resulting in low test results.
2. If there is precipitation in the dye solution before dyeing, it needs to be filtered to remove the precipitation before use.
3. There is a potential carcinogenic risk. For your safety and health, please wear a lab coat and disposable gloves for operation.
References:
[1]. Warren Strober.Trypan Blue Exclusion Test of Cell Viability. 2015 Nov 2;111:A3.B.1-A3.B.3. doi: 10.1002/0471142735.ima03bs111.
| Cas No. | 72-57-1 | SDF | |
| Canonical SMILES | CC1=CC(C2=CC=C(/N=N/C3=C(S(=O)([O-])=O)C=C4C=C(S(=O)([O-])=O)C=C(N)C4=C3O)C(C)=C2)=CC=C1/N=N/C5=C(S(=O)([O-])=O)C=C6C=C(S(=O)([O-])=O)C=C(N)C6=C5O.[Na+].[Na+].[Na+].[Na+] | ||
| Formula | C34H24N6Na4O14S4 | M.Wt | 960.81 |
| Solubility | Water : 150 mg/mL (156.12 mM) | 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 | |||
|
1 mg | 5 mg | 10 mg |
| 1 mM | 1.0408 mL | 5.2039 mL | 10.4079 mL |
| 5 mM | 0.2082 mL | 1.0408 mL | 2.0816 mL |
| 10 mM | 0.1041 mL | 0.5204 mL | 1.0408 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 ddH2O, 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: >60.00%
- COA (Certificate Of Analysis)
- SDS (Safety Data Sheet)
- Datasheet
Average Rating: 5 (Based on Reviews and 11 reference(s) in Google Scholar.)
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