We directly dissolved L012 (Wako Pure Chemical Industries, Ltd., Osaka, Japan) and 1,10-phenanthroline (Sinopharm Co., Shanghai, China) in ultrapure water to prepare a 4 mM stock solution. Luminol (Sigma-Aldrich) and ABEI (TCI, Tokyo, Japan) were respectively dissolved in NaOH (pH 12) solution to prepare 4 mM stock solutions. Next, 2-imidazolidinetione, 2-methyl-3-thiosemicarbazide, and pyrogallic acid were purchased from Shanghai Acmec Biochemical Co., Ltd. (Shanghai, China). Catechin was purchased from Solarbio (Beijing, China). Dopamine was purchased from J&K Scientific (Beijing, China); DMPO was purchased from Dojindo Molecular Technologies, Inc. (Kumamoto, Japan); and fluorescein and rhodamine B were purchased from Sangon Biotech (Shanghai, China). H2O2 was prepared fresh daily from 30% (vol/vol) H2O2 (Xinke Electrochemical Reagent Factory, Bengbu, China). All other reagents were obtained from Sinopharm Co. (Shanghai, China), of analytical grade. Ultrapure water was prepared by a Milli-Q system (Millipore, France) and used throughout.
Fluorescein
Manufactured by Sangon
Sourced in Japan, China
Fluorescein is a fluorescent dye commonly used as a laboratory reagent. It has the ability to emit light when excited by a specific wavelength of light, making it useful for various applications in scientific research and analysis.
Automatically generated - may contain errors
Lab products found in correlation
2 protocols using fluorescein
1
Preparation of Luminescent Reagents
2
Native PAGE Analysis of Protein-DNA Interactions
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Protein–protein interactions were examined in 8% w/v native PAGE, which uses the same Tris-glycine gel system as a standard SDS-PAGE, but without SDS. Electrophoretic mobility shift assay (EMSA) is another form of native PAGE to examine protein–DNA interactions in the Tris-Borate-EDTA buffer system. A 25-mer DNA oligo (25-mer, GTT TGT TATAAAAG TGT TACAATAC) taken from the gtfC gene promoter of S. mutans was chemically synthesized and labeled with fluorescein (Sangon, China). Double-stranded DNA was produced by annealing two oligos at 95°C for 5 min and cooled down to RT. DNA oligo at 20 μM was mixed with WalR or DBD and incubated in a binding buffer (20 mM Tris-HCl, pH 7.5, 100 mM NaCl, 50 mM KCl, and 1 mM DTT) for 15 min at RT. WalK 196–450 at various molar ratios to WalR or DBD was directly added into the mixtures and incubated for another 15 min. Before loading onto the gels, 50% v/v glycerol was added and samples were analyzed on an 8% w/v gel, on ice, using 120 V for 1 h. The gel was then visualized by UV for DNA, and further stained by CBB for proteins.
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