Abstract
Curcumin, the principal curcuminoid in the food spice turmeric, is insoluble in water at room temperature. We have previously solubilized curcumin in water with the application of heat (100 °C) and found that this solubilized curcumin could be used as a gel protein stain. However, heat solubilization in water solubilized only a small fraction of the curcuminoid (1.5%), making it relatively expensive to stain one gel ($1.5–2 per gel). Heat/pressure solubilization (121 °C/15 psi) helps increase the solubility of curcumin in water to only 5%. Therefore, we ventured into increasing curcumin solubility further by using detergents (Tween 20, Tween 80, or Triton X-100) combined with heat/pressure treatment. Curcumin solubilized with Tween 20, Tween 80 or Triton X-100 stained proteins efficiently as before. We could dramatically increase the solubility of curcumin by solubilizing in detergents such that we had to dilute the solubilized curcumin 1:100 with water to stain proteins. However, we had to use the soluble curcumin (extracted with heat/pressure alone) straight without dilution. Thus, the amount of curcumin solubilized with detergents was at least 100-fold, with 0.5% Tween 80 being the most efficient. Staining can be carried out in 30 min, and sensitivity of staining is similar or better than that obtained with Coomassie Brilliant Blue (CBB). It is unnecessary to destain the gel, and excess curcumin can be discarded into the sink, unlike CBB. Curcumin solubilized in Tween 20 has been found to be stable for a period of over 4 years. Therefore, curcumin solubilized in Tween 80 or Triton X-100 can serve as an efficient nontoxic and environment-friendly protein stain. Tween 80 has been used in cough syrups to make a colloidal suspension, and also in foods. Solubilizing curcumin with Tween 80 would be a useful alternative to increase curcumin bioavailability in clinical studies.
This is a preview of subscription content, log in via an institution.
Buying options
Tax calculation will be finalised at checkout
Purchases are for personal use only
Learn about institutional subscriptionsSpringer Nature is developing a new tool to find and evaluate Protocols. Learn more
References
D’souza A, Scofield RH (2009) Protein stains to detect antigen on membranes. Methods Mol Biol 536:433–440
Kurien BT, Scofield RH (1998) Heat mediated quick Coomassie blue protein staining and destaining of SDS-PAGE gels. Indian J Biochem Biophys 35:385–389
Steinberg TH (2009) Protein gel staining methods: an introduction and overview. Methods Enzymol 463:541–563 Review
Jin LT, Hwang SY, Yoo GS et al (2006) A mass spectrometry compatible silver Staining method for protein incorporating a new silver sensitizer in sodium dodecyl sulfate-polyacrylamide electrophoresis gels. Proteomics 6:2334–2337
Jin LT, Hwang SY, Yoo GS et al (2004) Sensitive silver staining of protein in sodium dodecyl sulfate-polyacrylamide gels using an azo dye, calconcarboxylic acid, as a silver-ion sensitizer. Electrophoresis 25:2494–2500
Møller HJ, Heinegård D, Poulsen JH (1993) Combined alcian blue and silver staining of subnanogram quantities of proteoglycans and glycosaminoglycans in sodium dodecyl sulfate-polyacrylamide gels. Anal Biochem 209:169–175
Pal JK, Godbole D, Sharma K (2004) Staining of proteins on SDS Polyacrylamide gels and on nitrocellulose membranes by Alta, a colour used as a cosmetic. J Biochem Biophys Methods 61:339–347
Lin CY, Wang V, Shui HA et al (2009) A comprehensive evaluation of imidazole-zinc reverse stain for current proteomic researches. Proteomics 9:696–709
Aggarwal BB, Kumar A, Aggarwal MS et al (2004) Curcumin derived from turmeric (Curcuma longa): a spice for all seasons. In: Bagchi D, Preuss HG (eds) Phytochemicals in cancer chemoprevention. CRC Press, Boca Raton, FL, pp 349–387
Aggarwal BB, Sung B (2009) Pharmacological basis for the role of curcumin in chronic diseases: an age-old spice with modern targets. Trends Pharmacol Sci 30:85–94
Bright JJ (2007) Curcumin and autoimmune disease. Adv Exp Med Biol 595:425–451 Review
Rowe DL, Ozbay T, O'Regan RM et al (2009) Modulation of the BRCA1 protein and induction of apoptosis in triple negative breast cancer cell lines by the polyphenolic. Breast Cancer 3:61–75
Anand P, Sundaram C, Jhurani S et al (2008) Curcumin and cancer: an “old-age” disease with an “age-old” solution. Cancer Lett 267:133–164 Review
Kurien BT, Singh A, Matsumoto H et al (2007) Improving the solubility and pharmacological efficacy of curcumin by heat treatment. Assay Drug Dev Technol 5:567–576
Kurien BT, Scofield RH (2009) Curry spice curcumin and prostate cancer. Mol Nutr Food Res 53:939–940
Kurien BT (2009) Comment on curcumin attenuates acrylamide-induced cytotoxicity and genotoxicity in HepG2 cells by ROS scavenging. J Agric Food Chem 57:5644–5646
Kurien BT, Scofield RH (2009) Bubbling hookah smoke through heat-solubilized curcumin/turmeric and incorporation of the curry spice as an additive or filter in cigarettes to minimize tobacco smoke-related toxicants. Med Hypotheses 73:462–463
Kurien BT, Matsumoto H, Scofield RH (2017) Nutraceutical value of pure curcumin. Pharmacogn Mag 13(Suppl 1):S161–S163
Kurien BT, D’Souza A, Scofield RH (2010) Heat-solubilized curry spice curcumin inhibits antibody–antigen interaction in in vitro studies: a possible therapy to alleviate autoimmune disorders. Mol Nutr Food Res 54:1202–1209
Laemmli UK (1970) Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 227:680–685
Douglas GH (1997) Colloidal aspects of ice cream—a review. Int Dairy J 7:363–373
Ema M, Hara H, Matsumoto M, Hirata-Koizumi M, Hirose A, Kamata E (2008) Evaluation of developmental neurotoxicity of polysorbate 80 in rats. Reprod Toxicol 25:89–99
Kurien BT, Dorri Y, Scofield RH (2012) Spicy SDS-PAGE gels: curcumin/turmeric as an environment-friendly protein stain. Methods Mol Biol 869:567–578
Author information
Authors and Affiliations
Corresponding author
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2018 Springer Science+Business Media, LLC, part of Springer Nature
About this protocol
Cite this protocol
Kurien, B.T., Thomas, R., Payne, A., Scofield, R.H. (2018). Heat/Pressure Treatment with Detergents Significantly Increases Curcumin Solubility and Stability: Its Use as an Environment-Friendly Protein Gel Stain. In: Kurien, B., Scofield, R. (eds) Protein Gel Detection and Imaging. Methods in Molecular Biology, vol 1853. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-8745-0_27
Download citation
DOI: https://doi.org/10.1007/978-1-4939-8745-0_27
Published:
Publisher Name: Humana Press, New York, NY
Print ISBN: 978-1-4939-8744-3
Online ISBN: 978-1-4939-8745-0
eBook Packages: Springer Protocols