Research Article
Affordable and Simple Protocol for Immunofluorescence Staining of Insulin Secreting iGL Cell Line 2D and 3D
Abstract
Immunofluorescence staining is a widely used technique to visualize the localization of specific proteins in cells. This protocol provides an affordable and simple method for immunofluorescence staining of the Insulin-GLase (iGL) cell line in both 2D and 3D culture conditions. We describe cost-effective alternatives for reagents and materials without compromising the quality of the results, such as the use of Bovine Serum Albumin (BSA) 3% as a blocking buffer instead of normal serum. Immunofluorescence staining on the iGL cell line is a valuable technique for visualizing the distribution and localization of target biomolecules. With careful consideration of cost-effective strategies and an understanding of the unique characteristics of the iGL cell line, it is possible to achieve reliable and informative immunofluorescence results even with budget constraints.
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- Sirenko O, Mitlo T, Hesley J, Luke S, Owens W, Cromwell EF. High-Content Assays for Characterizing the Viability and Morphology of 3D Cancer Spheroid Cultures. ASSAY DRUG DEV TECHN. 2015;13(7):402–414
Year 2024,
Volume: 41 Issue: 3, 590 - 594, 30.09.2024
Abstract
References
- 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;7:1.
- Bhattacharyya D, Hammond AT, Glick BS. High-Quality Immunofluorescence of Cultured Cells. 2010,p. 403–410.
- Maity B, Sheff D, Fisher RA. Immunostaining. 2013, p. 81–105.
- Barzowska A, Pucelik B, Pustelny K, Matsuda A, Martyniak A, Stępniewski J, et al. DYRK1A Kinase Inhibitors Promote β-Cell Survival and Insulin Homeostasis. Cells. 2021;10(9):2263.
- Davies MN, O’Callaghan BL, Towle HC. Glucose Activates ChREBP by Increasing Its Rate of Nuclear Entry and Relieving Repression of Its Transcriptional Activity. JBC. 2008; 283(35):24029–24038.
- Nierode G, Kwon PS, Dordick JS, Kwon SJ. Cell-Based Assay Design for High-Content Screening of Drug Candidates. J MICROBIOL BIOTECHN. 2016; 26(2):213–225.
- Liang Y, Jetton TL, Zimmerman EC, Najafi H, Berner DK, Matschinsky FM, et al. Effects of Glucose on Insulin Secretion, Glucokinase Activity, and Transgene Expression in Transgenic Mouse Islets Containing an Upstream Glucokinase Promoter-Human Growth Hormone Fusion Gene. Diabetes. 1994;43(9):1138–1145.
- Damalanka VC, Han Z, Karmakar P, O’Donoghue AJ, La Greca F, Kim T, et al. Discovery of Selective Matriptase and Hepsin Serine Protease Inhibitors: Useful Chemical Tools for Cancer Cell Biology. J Med Chem. 2019;62(2):480–490.
- Tseng SC, Hatchell D, Tierney N, Huang AJ, Sun TT. Expression of specific keratin markers by rabbit corneal, conjunctival, and esophageal epithelia during vitamin A deficiency. JCB.1984;99(6):2279–2286.
- Riley JK, Carayannopoulos MO, Wyman AH, Chi M, Moley KH. Phosphatidylinositol 3-Kinase Activity Is Critical for Glucose Metabolism and Embryo Survival in Murine Blastocysts. JBC. 2006;281(9):6010–6019.
- Ningsih SS, Avissa R, Stujanna EN, Listyaningsih E, Yashiro T, Sukarya WS. Evaluation of morphology and viability of spheroid derived from Insulin-GLase cell line: A model system to understand Type 2 Diabetes Mellitus. JECM. 2021;38(3):211-215.
- Sirenko O, Mitlo T, Hesley J, Luke S, Owens W, Cromwell EF. High-Content Assays for Characterizing the Viability and Morphology of 3D Cancer Spheroid Cultures. ASSAY DRUG DEV TECHN. 2015;13(7):402–414
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Details
| Primary Language | English |
|---|---|
| Subjects | Medical Biotechnology (Other) |
| Journal Section | Research Article |
| Authors | |
| Publication Date | September 30, 2024 |
| Submission Date | March 27, 2024 |
| Acceptance Date | July 8, 2024 |
| Published in Issue | Year 2024 Volume: 41 Issue: 3 |
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