Resazurin  (Synonyms: Diazoresorcinol)

Resazurin (Diazoresorcinol) is a water-soluble, non-toxic, stable, membrane-permeable blue non-fluorescent dye (faintly fluorescent). Resazurin is used as a redox indicator, can be reduced to pink, highly fluorescent Resorufin (Ex=530-560 nm, Em=590 nm) in living cells. Resazurin can be used for the detection of cell viability, toxicity, proliferation, migration and invasion in cells (human, plant and animal, bacterial and fungal)^[1][2].

Resazurin (Diazoresorcinol) is commonly used to measure the viability of bacterial and eukaryotic cells through its reduction to the fluorescent product resorufin. In TSBc medium, no viable bacteria were detected 24 hours after inoculation of F. tularensis LVS in the presence of Resazurin at a concentration of 44 μM. Reducing the concentration of Resazurin to 4.4 μM still resulted in a 10-fold reduction in the survival of F. tularensis LVS compared with medium alone. Both Resazurin treatments significantly reduced the number of viable F. tularensis LVS bacteria within 22 hours. Following infection of HEK293 cells for 22 hours, Resazurin treatment significantly reduced the number of viable F. tularensis LVS bacteria^[3].

Guide (The following is our recommended protocol. This protocol is provided as a guideline only and should be modified according to your specific needs).
1. Solution Preparation^[2][3]
1.1 Stock Solution Preparation
Solvent: DMSO or ddH₂O
Concentration: 1 mg/mL (optimize according to the experiment).
Storage: Aliquot and store at -20°C or -80°C protected from light. Avoid repeated freeze-thaw cycles.
1.2 Working Solution Preparation
Dilute with PBS or serum-free culture medium to 1 μg/mL (optimize according to the experiment).
Note: The working solution should be prepared immediately before use and protected from light.

2. Cell Viability
1. Thaw the Resazurin solution in a 37°C water bath.
2. Plate cells in a 96-well plate and wash with PBS (avoid light exposure).
3. Remove the PBS wash solution, then add 500 μL of Resazurin solution (1 μg/mL).
4. Place the plate in an incubator for 30 minutes (incubation time depends on cell type and cell number).
5. Measure Resazurin fluorescence using a spectrophotometer (Ex=530-560 nm, Em=590 nm).

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

229.19

C₁₂H₇NO₄

550-82-3

590

530/560

O=C1C=CC2=[N+](C3=C(OC2=C1)C=C(C=C3)O)[O-]

Room temperature in continental US; may vary elsewhere.

Please store the product under the recommended conditions in the Certificate of Analysis.

• [1]. Silva FSG, et al. Determination of Metabolic Viability and Cell Mass Using a Tandem Resazurin/Sulforhodamine B Assay. Curr Protoc Toxicol. 2016 May 4;68:2.24.1-2.24.15.  [Content Brief]

  [2]. Rampersad SN. Multiple applications of Alamar Blue as an indicator of metabolic function and cellular health in cell viability bioassays. Sensors (Basel). 2012;12(9):12347-60.  [Content Brief]

  [3]. Schmitt DM, et al. The use of resazurin as a novel antimicrobial agent against Francisella tularensis. Front Cell Infect Microbiol. 2013 Dec 6;3:93.  [Content Brief]