Sulforhodamine B sodium salt (Acid Red 52) (Synonyms: Acid Red XB) |
カタログ番号GC30365
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スルホローダミン B ナトリウム塩 (Acid Red 52) は、レーザー誘起蛍光 (LIF) から培養細胞の細胞タンパク質の定量化まで幅広い用途を持つ蛍光色素です。
Products are for research use only. Not for human use. We do not sell to patients.
Cas No.: 3520-42-1
Sample solution is provided at 25 µL, 10mM.
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Sulforhodamine B sodium salt (Acid Red 52) is a water-soluble red fluorescent/colorimetric dye with an anionic aminoxanthene structure, the maximum excitation/emission wavelength is 565nm/586nm. Under trichloroacetic acid fixation conditions, Sulforhodamine B sodium salt electrostatically binds to basic amino acid residues of intracellular proteins. After dissociation with alkaline solution, absorbance is measured to quantify total protein, thereby reflecting cell number by protein amount. Sulforhodamine B sodium salt can serve as a membrane-impermeant polar tracer. Sulforhodamine B sodium salt can be used in studies on adherent tumor cell proliferation assay, chemotherapeutic drug and small molecule cytotoxicity screening, and NCI large-scale antitumor drug screening[1-4].
References:[1] Rezaee M, Oskuee RK, Nassirli H, et al. Progress in the development of lipopolyplexes as efficient non-viral gene delivery systems. J Control Release. 2016 Aug 28;236:1-14.
[2] Shin CI, Kim M, Kim YC. Delivery of Niacinamide to the Skin Using Microneedle-Like Particles. Pharmaceutics. 2019 Jul 11;11(7):326.
[3] Chen H, Wang J, Zhang J, et al. L1EPO, a novel podophyllotoxin derivative overcomes P-glycoprotein-mediated multidrug resistance in K562/A02 cell line. Biol Pharm Bull. 2009 Apr;32(4):609-13.
[4] Maurice D, Singh T. A permeability test for acute corneal toxicity. Toxicol Lett. 1986 May;31(2):125-30.
Staining method of Sulforhodamine B sodium salt[1]
This protocol provides only a guideline. Please modify it according to specific needs.
1. Preparation of staining solution:
(1) Prepare the working solution: Dissolve Sulforhodamine B sodium salt in sterile water or PBS to make a 0.04% (wt/vol) working solution. Filtered through a 0.22μm membrane. Store any unused solution at -20°C or -80°C, in aliquots, and keep Sulforhodamine B sodium salt away from light.
2. Staining of adherent cells
(1) Cell seeding: seed cells in 96-well or 384-well culture plates. Add appropriate volume of growth medium. Incubate at 37°C in a humidified incubator with 5% CO2 until desired confluency (usually 70-80%).
(2) Cell treatment: add treatment solution (e.g., compound or transfection complex) to wells. Co-incubate with cells for appropriate time (e.g., 5 days).
(3) Cell fixation: directly add cold trichloroacetic acid (TCA) to each well to achieve a final concentration of 10-15% TCA. Incubate at 4°C for 1h. Then wash the plate by submerging in slow-running tap water 4 times. Remove excess water and air-dry at room temperature. Avoid direct water flow impacting the well bottom to prevent cell monolayer detachment. Ensure the plate is completely dry before proceeding to the next step.
(4) Staining: add 0.04% (wt/vol) Sulforhodamine B sodium salt solution to each well. Incubate at room temperature for 1h.
(5) Wash away unbound dye: quickly wash 4 times with 1% (vol/vol) acetic acid to remove unbound dye. Air-dry at room temperature. Washing must be rapid and uniform. Avoid bubbles hindering washing. Dried plates can be stored long-term at room temperature.
3. Detection
(1) Dissolution of bound dye: add 10mM Tris base solution (pH=10.5) to each well. Shake on an orbital shaker for 10min to dissolve protein-bound dye.
(2) Absorbance measurement: measure absorbance at 510nm using a microplate reader. Subtract background absorbance (wells containing medium only) before measurement. Ensure absorbance values are within the linear range.
Precautions:
(1) It is recommended to set untreated control group and blank control group (medium only) for calculating cell growth inhibition rate.
(2) Sulforhodamine B sodium salt is a colorimetric dye and does not require protection from light, but attention should be paid to washing uniformity to avoid inter-well variation.
(3) For safety and health, wear lab coat and disposable gloves during operation. Trichloroacetic acid is corrosive and should be handled with care in a fume hood.
References:[1] Orellana EA, Kasinski AL. Sulforhodamine B (SRB) Assay in Cell Culture to Investigate Cell Proliferation. Bio Protoc. 2016 Nov 5;6(21):e1984.
| Cas No. | 3520-42-1 | SDF | |
| 同義語 | Acid Red XB | ||
| Canonical SMILES | O=S(C1=CC=C(C2=C3C=CC(N(CC)CC)=CC3=[O+]C4=C2C=CC(N(CC)CC)=C4)C(S(=O)([O-])=O)=C1)([O-])=O.[Na+] | ||
| Formula | C27H29N2NaO7S2 | M.Wt | 580.65 |
| 溶解度 | DMSO : 50 mg/mL (86.11 mM);Water : 20 mg/mL (34.44 mM) | Storage | Store at -20°C |
| 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 | |||
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1 mg | 5 mg | 10 mg |
| 1 mM | 1.7222 mL | 8.611 mL | 17.2221 mL |
| 5 mM | 344.4 μL | 1.7222 mL | 3.4444 mL |
| 10 mM | 172.2 μL | 861.1 μL | 1.7222 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: >98.50% Appearance: A solid
- COA (Certificate of Analysis)
- SDS (Safety Data Sheet)
- Datasheet
Average Rating: 5 (Based on Reviews and 4 reference(s) in Google Scholar.)