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Sulforhodamine B sodium salt Dyes chemical

Cat.No.S5976

Sulforhodamine B (SRB, Acid Red 52, Kiton Red 620) sodium salt is a protein-specific fluorescent dye that is utilized to quantitate cell expansion, growth, and migration.
Sulforhodamine B sodium salt Dyes chemical Chemical Structure

Chemical Structure

Molecular Weight: 580.65

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Quality Control

Batch: S597601 Water]100 mg/mL]false]DMSO]Insoluble]false]Ethanol]Insoluble]false Purity: 99.03%
99.03

Solubility

In vitro
Batch:

Water : 100 mg/mL

DMSO : Insoluble
(Moisture-contaminated DMSO may reduce solubility. Use fresh, anhydrous DMSO.)

Ethanol : Insoluble

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In vivo
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In vivo Formulation Calculator (Clear solution)

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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.

Chemical Information, Storage & Stability

Molecular Weight 580.65 Formula
C27H29N2NaO7S2
 Storage (From the date of receipt) 3 years -20°C powder
CAS No. 3520-42-1 -- Storage of Stock Solutions

Synonyms SRB, Acid Red 52, Kiton Red 620 Smiles [Na+].CCN(CC)C1=CC2=C(C=C1)C(=C3C=CC(C=C3O2)=[N+](CC)CC)C4=C(C=C(C=C4)[S]([O-])(=O)=O)[S]([O-])(=O)=O

Mechanism of Action

In vitro

Sulforhodamine B colorimetric assay in cell culture to analyze cell proliferation
1. Prepare treatment solutions with sufficient volume for triplicates in 96-well plates (50 μl per replicate) or six replicates in 384-well plates (10 μl per replicate). Ensure volumes account for pipetting variations.
2. Treatment solutions can be prepared in either aqueous solution (e.g., Opti-MEM for transfections) or a suitable solvent (e.g., DMSO).
3. Remove the medium from cell monolayers and wash cells once with sterilized PBS. Add 1 ml (for 100 mm plates) of 0.25% trypsin to cover the cell growth surface evenly.
4. Incubate at 37 °C for 5 minutes or until cells start to dissociate. Inactivate trypsin with 10 volumes of culture medium containing FBS. Mix up and down to obtain a homogeneous single-cell suspension.
5. Transfer the cell suspension to a sterile Falcon tube.
6. Determine cell concentration using a hematocytometer chamber with a 1:1 mixture of cell suspension and 0.4% trypan blue solution. Optionally, spin down cells before counting to wash trypsin and resuspend in a growth medium.
7. Adjust cell concentration with growth medium (10% FBS) for appropriate cell seeding density per well.
8. Mix treatment solutions and dispense 50 μl (96-well format) or 10 μl (384-well format) into each well.
9. Thoroughly mix the cell suspension and add 50 μl (96-well format) or 10 μl (384-well format) to each well with treatment solutions.
Note: Ensure even cell distribution in the well bottom, avoiding shaking to prevent 'ring effects.
10. Set aside three wells for untreated or vehicle control and three wells for background subtraction and incubate the plate at 37 °C with 5% CO2 until ready for reading.
11. Add 25 μl (96-well format) or 5 μl (384-well format) of cold 50% TCA directly to medium supernatant. Incubate plates at 4 °C for 1 hour without mixing.
12. Wash plates four times by submerging in slow-running tap water, and tapping excess water into a paper towel. Allow the plate to air-dry at room temperature.
13. Add 50 μl (96-well format) or 20 μl (384-well format) of 0.04% SRB solution to each well.
14. incubate at room temperature for 1 hour and Rinse plates four times with 1% acetic acid (200 μl for 96-well format or 30 μl for 384-well format). Allow the plate to air-dry at room temperature.
15. Add 50 μl to 100 μl of 10 mM tris base solution (pH 10.5) to each well and shake the plate on an orbital shaker for 10 minutes to solubilize the protein-bound dye.
16. Measure the absorbance at 510 nm in a microplate reader.
References

Clinical Trial Information

(data from https://clinicaltrials.gov, updated on 2024-05-22)

NCT Number Recruitment Conditions Sponsor/Collaborators Start Date Phases
NCT04862260 Recruiting
Pancreatic Ductal Adenocarcinoma|Pancreatic Cancer|Pancreas Cancer|Metastatic Cancer
CHU de Quebec-Universite Laval|Canadian Institutes of Health Research (CIHR)|Biovalorem
 October 4 2021 Early Phase 1
NCT03451513 Completed
Eating Disorder|Body Image
San Diego State University|National Institute on Minority Health and Health Disparities (NIMHD)
 January 24 2018 Not Applicable

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