DISCOVERIES (ISSN 2359-7232), 2025, volume 13
ORIGINAL ARTICLE
CITATION: Shahid M, Iftikhar K, Simjee SU. Impact of Triton X-100 on Notch 1 Surface Receptor Immunofluorescence: A Cautionary Study. Discoveries 2025, 13(1): e206. DOI: 10.15190/d.2025.5
Impact of Triton X-100 on Notch 1 Surface Receptor Immunofluorescence: A Cautionary Study
Maha Shahid 1,#, Kanwal Iftikhar 1 # , Shabana Usman Simjee 1,2,*
- 1 Hussain Ebrahim Jamal Research Institute of Chemistry, International Center for Chemical and Biological Sciences, University of Karachi, Karachi 75270, Pakistan
- 2 Dr. Panjwani Center for Molecular Medicine and Drug Research, International Center for Chemical and Biological Sciences, University of Karachi, Karachi 75270, Pakistan
- #Contributed equally to this work.
* Corresponding author: Shabana Usman Simjee, H.E.J. Research Institute of Chemistry, International Center for Chemical and Biological Sciences, University of Karachi, Karachi 75270, Pakistan; Email: shabana.simjee@iccs.edu.
Abstract
Triton X-100 is a cell-permeabilizing agent, widely used in molecular studies to enhance the permeability of cellular membranes. It has detrimental effects on the cell surface receptors thereby not suitable for surface receptor protein studies. Therefore, in the present study the protein expression of Notch surface receptor was analyzed with and without exposure of cell surfactant i.e. Triton X-100. It was observed that cultured cortical cells treated with Triton X-100 gave false protein expression due to the disruption of cellular membrane. On the contrary, cells without surfactant treatment exhibit fluorescence proportional to the true presence of Notch 1 receptors. This method can be followed for the analysis of any surface receptor protein through immunofluorescence to exclude any false expression.
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REFERENCES
1. Kopan R. Notch Signaling. Cold Spring Harb Perspect Biol 2012; 4(10). doi: 10.1101/CSHPERSPECT.A011213.
2. Wu L, Sun T, Kobayashi K, Gao P, Griffin JD. Identification of a Family of Mastermind-Like Transcriptional Coactivators for Mammalian Notch Receptors. Mol Cell Biol 2002; 22(21), 7688–7700. doi.:10.1128/MCB.22.21.7688-7700.2002/ASSET/CE456B56-BDDC-4130-89DF-E547D1153F48/ASSETS/GRAPHIC/MB2120406010.JPE
3. Jamur MC, Oliver C. Permeabilization of cell membranes. Immunocytochemical methods and protocols. 2010:63-6. doi: 10.1007/978-1-59745-324-0_9
4. Benoit J, Cormier M, Wepierre J. Comparative effects of four surfactants on growth, contraction and adhesion of cultured human fibroblasts. Cell Biol Toxicol 1988; 4:1, 4(1), 111–122. doi: 10.1007/BF00141290.
5. Borner MM, Schneider E, Pirnia F, Sartor O, Trepel JB, Myers CE. The detergent Triton X-100 induces a death pattern in human carcinoma cell lines that resembles cytotoxic lymphocyte-induced apoptosis. FEBS Letters 1994; 353, 129–132. doi: 10.1016/0014-5793(94)01023-4.
6. Laouar L, Lowe KC, Mulligan BJ. Yeast responses to nonionic surfactants. Enzyme Microb Technol 1996; 18(6), 433–438. doi: 10.1016/0141-0229(95)00122-0.
7. Dayeh VR, Chow SL, Schirmer K, Lynn DH, Bols NC. Evaluating the toxicity of Triton X-100 to protozoan, fish, and mammalian cells using fluorescent dyes as indicators of cell viability. Ecotoxicol Environ Saf 2004; 57(3), 375–382. doi: 10.1016/S0147-6513(03)00083-6.
8. Giordano G, Costa LG. Primary neurons in culture and neuronal cell lines for in vitro neurotoxicological studies. Methods Mol Biol 2011; 758, 13–27. doi: 10.1007/978-1-61779-170-3_2.
9. Vernay A, Cosson P. Immunofluorescence labeling of cell surface antigens in Dictyostelium. BMC research notes. 2013 Dec;6:1-4. doi: 10.1186/1756-0500-6-317.
10. Cheng R, Zhang F, Li M, Wo X, Su YW, Wang W. Influence of Fixation and Permeabilization on the Mass Density of Single Cells: A Surface Plasmon Resonance Imaging Study. Front Chem 2019; 7, 588. doi: 10.3389/fchem.2019.00588.
11. Koley, D., & Bard, A. J. Triton X-100 concentration effects on membrane permeability of a single HeLa cell by scanning electrochemical microscopy (SECM). Proc Natl Acad Sci U S A 2010; 107(39), 16783–16787. doi: 10.1073/pnas.1011614107.
