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  3. Diphenyl Blue

Diphenyl Blue  (Synonyms: Direct Blue 14; Trypan Blue)

Cat. No.: HY-D0970
COA Handling Instructions

Diphenyl Blue (Trypan Blue) is a cell active dye, the most commonly used dye for the identification of dead cells, of en used to test cell membrane integrity and cell viability. Diphenyl Blue staining is one of the methods for tissue and cell culture. When cells are deactivated or have incomplete cell membranes, Diphenyl Blue can stain them Blue. Normal living cells with intact cell membranes reject Diphenyl blue and do not stain them blue. However, macrophages are capable of phagocytosis of Diphenyl Blue, so it can be used as a living stain for macrophages.

For research use only. We do not sell to patients.

Diphenyl Blue Chemical Structure

Diphenyl Blue Chemical Structure

CAS No. : 72-57-1

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Customer Review

Based on 2 publication(s) in Google Scholar

Top Publications Citing Use of Products
  • Biological Activity

  • Purity & Documentation

  • References

  • Customer Review

Description

Diphenyl Blue (Trypan Blue) is a cell active dye, the most commonly used dye for the identification of dead cells, of en used to test cell membrane integrity and cell viability. Diphenyl Blue staining is one of the methods for tissue and cell culture. When cells are deactivated or have incomplete cell membranes, Diphenyl Blue can stain them Blue. Normal living cells with intact cell membranes reject Diphenyl blue and do not stain them blue. However, macrophages are capable of phagocytosis of Diphenyl Blue, so it can be used as a living stain for macrophages[1].

In Vitro

Preparation of Trypan Blue working solution
1.1 Preparation of the stock solution
Dissolve 10 mg of Trypan Blue in 100 mL 0.85% Nacl to obtain 0.4% Trypan Blue.
Note: It is recommended to store the stock solution at -20℃ -80℃ away from light and avoid repetitive freeze-thaw cycles.
1.2 Preparation of Trypan Blue working solution
Dilute the stock solution in 0.85% Nacl to obtain 0.04% of Trypan Blue working solution.
Note: Please adjust the concentration of Trypan Blue working solution according to the actual situation.
Cell staining
2.1 For suspension cells, centrifuge at 1000 g at 4℃ for 3-5 minutes and then discard the supernatant. Wash twice with PBS, 5 minutes each time.
For adherent cells, discard the cell culture medium, and add trypsin to dissociate cells to make a single-cell suspension. Centrifuge at 1000 g at 4℃ for 3-5 minutes and then discard the supernatant. Wash twice with PBS, 5 minutes each time.
2.2 Add 1 mL of rypan Blue working solution, and then incubate at room temperature for 5 minutes.
2.3 Centrifuge at 400 g at 4℃ for 3-4 minutes and then discard the supernatant.
2.4 Wash twice with PBS, 5 minutes each time.
2.5 Resuspend cells with serum-free cell culture medium or PBS, and then detect by fluorescence microscope.

MedChemExpress (MCE) has not independently confirmed the accuracy of these methods. They are for reference only.

Molecular Weight

960.81

Formula

C34H24N6Na4O14S4

CAS No.
Appearance

Solid

Color

Light brown to black

SMILES

CC1=CC(C2=CC=C(/N=N/C3=C(S(=O)(O[Na])=O)C=C4C=C(S(=O)(O[Na])=O)C=C(N)C4=C3O)C(C)=C2)=CC=C1/N=N/C5=C(S(=O)(O[Na])=O)C=C6C=C(S(=O)(O[Na])=O)C=C(N)C6=C5O

Shipping

Room temperature in continental US; may vary elsewhere.

Storage

4°C, sealed storage, away from moisture and light

*In solvent : -80°C, 6 months; -20°C, 1 month (sealed storage, away from moisture and light)

Solvent & Solubility
In Vitro: 

DMSO : 25 mg/mL (26.02 mM; Need ultrasonic; Hygroscopic DMSO has a significant impact on the solubility of product, please use newly opened DMSO)

H2O : 1 mg/mL (1.04 mM; ultrasonic and warming and heat to 60°C)

Preparing
Stock Solutions
Concentration Solvent Mass 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
View the Complete Stock Solution Preparation Table

* Please refer to the solubility information to select the appropriate solvent. Once prepared, please aliquot and store the solution to prevent product inactivation from repeated freeze-thaw cycles.
Storage method and period of stock solution: -80°C, 6 months; -20°C, 1 month (sealed storage, away from moisture and light). When stored at -80°C, please use it within 6 months. When stored at -20°C, please use it within 1 month.

* Note: If you choose water as the stock solution, please dilute it to the working solution, then filter and sterilize it with a 0.22 μm filter before use.

  • Molarity Calculator

  • Dilution Calculator

Mass (g) = Concentration (mol/L) × Volume (L) × Molecular Weight (g/mol)

Mass
=
Concentration
×
Volume
×
Molecular Weight *

Concentration (start) × Volume (start) = Concentration (final) × Volume (final)

This equation is commonly abbreviated as: C1V1 = C2V2

Concentration (start)

C1

×
Volume (start)

V1

=
Concentration (final)

C2

×
Volume (final)

V2

In Vivo Dissolution Calculator
Please enter the basic information of animal experiments:

Dosage

mg/kg

Animal weight
(per animal)

g

Dosing volume
(per animal)

μL

Number of animals

Recommended: Prepare an additional quantity of animals to account for potential losses during experiments.
Calculation results:
Working solution concentration: mg/mL
Purity & Documentation

References

Complete Stock Solution Preparation Table

* Please refer to the solubility information to select the appropriate solvent. Once prepared, please aliquot and store the solution to prevent product inactivation from repeated freeze-thaw cycles.
Storage method and period of stock solution: -80°C, 6 months; -20°C, 1 month (sealed storage, away from moisture and light). When stored at -80°C, please use it within 6 months. When stored at -20°C, please use it within 1 month.

Optional Solvent Concentration Solvent Mass 1 mg 5 mg 10 mg 25 mg
H2O / DMSO 1 mM 1.0408 mL 5.2039 mL 10.4079 mL 26.0197 mL
DMSO 5 mM 0.2082 mL 1.0408 mL 2.0816 mL 5.2039 mL
10 mM 0.1041 mL 0.5204 mL 1.0408 mL 2.6020 mL
15 mM 0.0694 mL 0.3469 mL 0.6939 mL 1.7346 mL
20 mM 0.0520 mL 0.2602 mL 0.5204 mL 1.3010 mL
25 mM 0.0416 mL 0.2082 mL 0.4163 mL 1.0408 mL

* Note: If you choose water as the stock solution, please dilute it to the working solution, then filter and sterilize it with a 0.22 μm filter before use.

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Help & FAQs
  • Do most proteins show cross-species activity?

    Species cross-reactivity must be investigated individually for each product. Many human cytokines will produce a nice response in mouse cell lines, and many mouse proteins will show activity on human cells. Other proteins may have a lower specific activity when used in the opposite species.

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Diphenyl Blue
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