Rbitc

Manufactured by Merck Group

Sourced in United States

RBITC is a fluorescent dye used in various laboratory applications. It is a derivative of rhodamine with the chemical formula C 27 H 29 N 2 O 3 Cl. RBITC is commonly used for labeling and tracking biomolecules, such as proteins and cells, in research and diagnostic settings.

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Rhodamine B isothiocyanate (RBITC), (7 citations)

5 protocols using rbitc

Fluorescent Alginate Scaffold Synthesis

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For the purpose of window chamber intravital microscopy (IVM) experiments, we transplanted fluorescent scaffolds, in which the alginate (Sigma-Aldrich, USA) was labeled with rhodamine B isothiocyanate (RBITC) (Sigma Aldrich, Milwaukee, WI).
alginate was labeled with RBITC according to a method previously reported by Mladenovska et al. [24 (link)]. Briefly, an aqueous 2w/w% alginate solution was prepared and adjusted to a pH=8 by adding 1M sodium hydroxide (Anachemia, Reno, NV). An RBITC solution was prepared by mixing 1 mg of RBITC in 1 mL DMSO (Fisher, Fairlawn, NJ) followed by slow addition into the alginate solution. The alginate-RBITC mixture was stirred for 1 h at 40 °C. After stirring, 0.5349 g of NH₄Cl (Sigma Aldrich) was added and mixed until fully dissolved. The alginate-RBITC solution was dialyzed in darkness overnight. Water baths were frequently changed with distilled deionized water. Subsequently, the alginate-RBITC solution was poured into 50 mL polypropylene conical vials until approximately three-quarters full, flash frozen at −80°C and lyophilized until fully dry. Lyophilized alginate-RBITC was stored at −20 °C until use.

Ziv K., Nuhn H., Ben-Haim Y., Sasportas L.S., Kempen P.J., Niedringhaus T.P., Hrynyk M., Sinclair R., Barron A.E, & Gambhir S.S. (2014). A tunable silk-alginate hydrogel scaffold for stem cell culture and transplantation. Biomaterials, 35(12), 3736-3743.

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Fluorescent Labeling of βγ-CAT

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The labeling reactions were performed in accordance with a previously described procedure with some modifications^{52 (link)}. Purified βγ-CAT was dialyzed overnight in a cross-linking solution (NaHCO₃ 7.56 g, Na₂CO₃ 1.06 g, and NaCl 7.36 g dissolved in 1 L of water; pH 9.0) using a 3,500 Da dialysis membrane (Biosharp, Cat BS-QT-021), and 1 mg of RBITC (Sigma, Cat 283924) was dissolved in 1 mL of DMSO (Sigma, Cat D2650). The RBITC solution was slowly added to βγ-CAT to give βγ-CAT: RBITC = 1 mg:150 μg and the sample was incubated overnight at 4 °C, protected from light. NH₄Cl (50 mM) was added and the solution was incubated at 4 °C for 2 hours to terminate the reaction. The dialyzed samples were concentrated to 1 mg mL⁻¹ with PBS and stored at 4 °C in the dark.

Zhao Z., Shi Z.H., Ye C.J, & Zhang Y. (2022). A pore-forming protein drives macropinocytosis to facilitate toad water maintaining. Communications Biology, 5, 730.

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Synthesis of RBITC-labeled LMCC

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A total of 0.5 g LMCC and 0.5 g RBITC (Sigma-Aldrich; Merck KGaA) were dissolved in 10 ml PBS (pH 7.4). Following stirring for 24 h in the dark, the reaction solution was transferred into the dialysis bag (MWCO 3500) and dialyzed against water (6 times, 2 l per time) for 48 h in the dark. The solution in the dialysis bag was freeze dried by a lyophilizer (FD-1-50; Biocool, Ltd., Beijing, China) for 24 h and RBITC-labeled LMCC (RBITC-LMCC) was obtained as a red powder (27 (link)).

Kou C.H., Han J., Han X.L., Zhuang H.J, & Zhao Z.M. (2017). Preparation and characterization of the Adriamycin-loaded amphiphilic chitosan nanoparticles and their application in the treatment of liver cancer. Oncology Letters, 14(6), 7833-7841.

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Biodistribution and Excretion Analysis in Zebrafish

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PP pellets,
xylene, THF, and RBITC were purchased from Sigma-Aldrich. Ethyl alcohol
(anhydrous, 99.9%) was purchased from Samchun Chemicals (Seoul, Korea).
Polytetrafluoroethylene (PTFE, 0.2 μm pore size, 47 mm, Omnipore
hydrophilic membrane filter) and polyvinylidene fluoride (PVDF, 0.2
μm pore size, 47 mm, Durapore hydrophilic membrane filter) were
purchased from Millipore, USA. To observe the biodistribution and
excretion using ZFEs, pronase (P8811) and tricaine (ethyl 3-aminobenzoate
methanesulfonate salt) were purchased from Sigma-Aldrich. EW was prepared
by dissolving 5 mM NaCl, 0.17 mM KCl, 0.33 mM CaCl₂, and
0.33 mM MgSO₄ in 1 L of DW according to previous work.^{40 (link)}

Lee W.S., Kim H., Sim Y., Kang T, & Jeong J. (2022). Fluorescent Polypropylene Nanoplastics for Studying Uptake, Biodistribution, and Excretion in Zebrafish Embryos. ACS Omega, 7(2), 2467-2473.

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Synthesis of Fluorescent Silica Nanoparticles

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Tetraethyl orthosilicate (TEOS), NH₄OH (aqueous solution, 28%), RBITC, and 3-aminopropyltriethoxysilane (APTES) were all purchased from Sigma-Aldrich Co. (St. Louis, MO, USA). Ethanol (EtOH) anhydrous was purchased from Duksan, Inc. (Ansan, Korea).

Lee A.R., Nam K., Lee B.J., Lee S.W., Baek S.M., Bang J.S., Choi S.K., Park S.J., Kim T.H., Jeong K.S., Lee D.Y, & Park J.K. (2019). Hepatic Cellular Distribution of Silica Nanoparticles by Surface Energy Modification. International Journal of Molecular Sciences, 20(15), 3812.

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