Baicalin (purity ≥ 98%) (Wako, Osaka, Japan) was dissolved in methanol (Wako) and added into the medium to a final concentration of 0, 0.01, 0.1, and 1 μM. To determine the optimal concentration for obtaining the optimal effect of Baicalin and the toxicity of Baicalin to HCEM cells, different concentrations (0.01, 0.1, and 1 μM) of Baicalin were used.
Baicalin
Manufactured by Fujifilm
Sourced in Japan, United States
Baicalin is a laboratory reagent used for various analytical and research applications. It is a natural compound derived from the roots of the Scutellaria baicalensis plant. Baicalin exhibits characteristic absorbance and fluorescence properties that can be utilized in spectrophotometric and fluorometric assays. The core function of Baicalin is to serve as a versatile analytical tool in research and testing environments.
Automatically generated - may contain errors
Lab products found in correlation
Geniposide, by Fujifilm (2 mentions)
6 well plate, by BD (2 mentions)
Methanol, by Fujifilm (2 mentions)
Baicalein, by Fujifilm (2 mentions)
Glabridin, by Fujifilm (1 mentions)
Naringin, by Merck Group (1 mentions)
Baicalein, by Merck Group (1 mentions)
Gallic acid, by Merck Group (1 mentions)
Nodakenin, by ChemFaces (1 mentions)
Cimifugin, by ChemFaces (1 mentions)
Wogonin, by ChemFaces (1 mentions)
Anti brdu antibody, by Roche (1 mentions)
96 well plate, by BD (1 mentions)
Varioscan flash, by Thermo Fisher Scientific (1 mentions)
Bromodeoxyuridine (brdu), by Roche (1 mentions)
Revertra ace first strand cdna synthesis kit, by Toyobo (1 mentions)
Ethylene diamine tetraacetic acid (edta), by Fujifilm (1 mentions)
Rneasy mini kit, by Qiagen (1 mentions)
Sybr green realtime pcr master mix, by Toyobo (1 mentions)
Lightcycler 480, by Roche (1 mentions)
11 protocols using baicalin
1
Optimization of Baicalin Concentration for HCEM Cells
2
Standardized Extraction of Herbal Compounds
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All herbal drugs used in this study were purchased from Tochimoto Tenkaido (Osaka, Japan). The Polygala Root extract (ext.), Areca ext., and Quercus Bark ext. were prepared from Polygala Root (TMPW28828), Areca (TMPW28896), and Quercus Bark (TMPW28902), respectively, using following procedure. Each herbal drug (30.0 g) was boiled gently with 300 mL of purified water for 60 min. Then, the supernatant of the decoction was lyophilized to obtain dry extract. The extracts were dissolved in purified water at a concentration of 10 mg/mL and kept at − 20 °C prior to use. The voucher specimen of these herbal drug extracts were deposited in the Museum of Materia Medica, Institute of Traditional Medicine (TMPW), University of Toyama, Japan. The natural compounds baicalin and glabridin were purchased from FUJIFILM Wako pure chemical corporation (Osaka, Japan). The structure of baicalin and glabridin were described (see the Electric Supplementary Material).
3
HPLC Analysis of Herbal Biomarkers
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The reference standard components, gallic acid (97.5–102.5%), naringin (95.0%), and baicalein (98.0%) were purchased from Merck KGaA (Darmstadt, Germany); cimifugin (98.0%), nodakenin (99.5%), oxypeucedanin hydrate (98.0%), and wogonin (98.9%) were purchased from ChemFaces Biochemical Co., Ltd. (Wuhan, China); narirutin (99.5%), glycyrrhizin (99.1%), and pulegone (99.3%) were purchased from Biopurify Phytochemicals (Chengdu, China); geniposide (≥98.0%) and baicalin (98.0%) were purchased from Wako Chemicals (Osaka, Japan); paeoniflorin (99.4%), ferulic acid (98.0%), neohesperidin (98.4%), arctiin (98.1%), wogonoside (98.9%), and arctigenin (99.4%) were purchased from Shanghai Sunny Biotech (Shanghai, China); and decursin (98.0%) and decursinol angelate (98.0%) were purchased from NPC Bio Technology (Yeongi, Korea). The chemical structures of these reference compounds are shown Fig. 1 . HPLC-grade solvents (methanol, acetonitrile, and water) and ACS reagent-grade formic acid (≥98.0%) for test solution preparation and simultaneous analysis were purchased from J. T. Baker (Phillipsburg, NJ, USA) and Merck KGaA (Darmstadt, Germany), respectively.![]()
Chemical structures of the 20 biomarker components in HYT.
4
Baicalin effects on neuropathic pain
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Baicalin was purchased from Wako (Wako Pure Chemical Industries, Osaka, Japan) and dissolved in 2% DMSO and saline. The stock solution was filtered through a 0.22 μm membrane filter before use. For behavioural experiments, rats were randomly divided into four groups (10 rats in each group): a sham group, a CCI group, a CCI group treated with Baicalin (15 μg/Kg) and a CCI group treated with Baicalin (30 μg/Kg). Rat received injection (i.p.) of Baicalin daily at 24 hr intervals, while sham and CCI animals received the same volume of DMSO in saline.
5
Baicalin Effects on HCEM Cell Proliferation
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HCEM cells (2.0 × 103 cells per well) were seeded in 96-well plates (FALCON). After reaching 60% confluence, there was a stepwise reduction of FBS (Daiichi Chemical) levels in the medium to 0%. Subsequently, 0–1 μM baicalin (Wako) was added and after 24 h, the cellular proliferative potential was assessed using the Cell proliferation ELISA 5-brom-2-deoxyuridine (BrdU) Kit (Roche Diagnostics, Basel, Switzerland). Each well was supplemented with BrdU (Roche Diagnostics) and cultured for 2 h at 37 °C, 5% CO2 gas-phase conditions. Next, each well was washed and treated with the anti-BrdU antibody (Roche Diagnostics). Moreover, we measured the absorbance of the developed color using a microplate reader (Varioscan Flash; Thermo Fisher Scientific, Waltham, MA, USA) at 370 nm.
6
Baicalin Modulates Osteoclastogenesis Markers
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HCEM cells (1.0 × 105 cells per well) were seeded in 6-well plates (FALCON). After reaching confluence, the FBS (Daiichi Chemical) concentration in the medium underwent stepwise reduction to 0%. Subsequently, 0–1 μM baicalin (Wako) was added, and after 48 h, the cells were harvested with trypsin (Nacalai Tesque)/EDTA (Wako) treatment. Furthermore, total ribonucleic acid (RNA) was harvested using the RNeasy Mini Kit (Qiagen, Venlo, Netherlands) and quantified by measuring the absorbance at 260 nm using a spectrophotometer BioSpec-nano (Shimadzu Corporation, Kyoto, Japan). Next, 1 μg of purified total RNA was reverse-transcribed into complementary deoxyribonucleic acid (cDNA) using a ReverTra Ace first-strand cDNA synthesis kit (Toyobo, Osaka, Japan). Furthermore, we analyzed OPG and RANKL gene expression levels using Light Cycler 480® (Roche Diagnostics) and SYBR Green Real-time PCR Master Mix (Toyobo). Table 1 shows the primer sequences.Table 1
Primer sequence.
| Gene | Sequence (5′→3′) |
| OPG | Forward: CTG CTG AAG CTG TGG AAA |
| RANKL | Forward: CAC ACC TCA CCA TCA ATG |
| GAPDH | Forward: ATG GCC TTC CGT GTT CCT |
7
Dkk-1 and Baicalin Regulate HCEM Cells
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HCEM cells (1.0 × 105 cells per well) were seeded in 6-well plates and cultured in 10% FBS (Daiichi Chemical)-containing α-MEM (Sigma Aldrich). After reaching confluency, Dkk-1 (R&D system) and baicalin (Wako) were added at 100 ng/mL and 0–1 μM, respectively. Protein extraction and Western blot analysis were performed 7 days after baicalin addition as mentioned earlier, using the same blocking agents and antibodies.
8
Evaluation of Anticancer Compounds
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Dimethyl sulfoxide (DMSO), sodium dodecyl sulfate (SDS), baicalin, baicalein, and cisplatin were purchased from Wako Pure Chemical Industries, Ltd. (Osaka, Japan). Dulbecco's modified Eagle's medium (DMEM) was purchased from Invitrogen (Carlsbad, CA, USA), and fetal bovine serum (FBS) was purchased from Nichirei Bioscience (Tokyo, Japan). Primary antibodies against CD44 and cPARP were purchased from Cell Signaling Technology (Danvers, MA, USA), and primary antibodies against phospho-CHK1 (S301) and β-actin were purchased from Sigma-Aldrich (St. Louis, MO, USA). The horseradish peroxidase-conjugated secondary anti-mouse immunoglobulin G (IgG) and anti-rabbit IgG antibodies were purchased from Santa Cruz Biotechnology, Inc. (Santa Cruz, CA, USA). The protein assay kit was purchased from Bio-Rad (Herndon, VA, USA). Liquid chromatography-grade acetonitrile, acetic acid, ethyl acetate, methanol, 1-butanol, 2-propanol, baicalein, baicalin, and trifluoroacetic acid [for use in the high-performance liquid chromatography (HPLC) experiments described below] were purchased from Wako Pure Chemical Industries, Ltd. Milli-Q plus water (Millipore, Bedford, MA, USA) was used in the present study. All other chemicals were purchased from Wako Pure Chemical Industries, Ltd., except where otherwise noted.
9
Keigairengyoto: Herbal Extract Antimicrobial Evaluation
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Keigairengyoto was supplied by Tsumura & Co. (lot number 2140050010, Tokyo, Japan) as a powdered extract obtained by spray-drying a hot water extract mixture of the following seventeen crude drugs: Scutellariae radix, Phellodendri cortex, Coptidis rhizoma, Platycodi radix, Aurantii fructus immaturus, Schizonepetae spica, Bupleuri radix, Gardeniae fructus, Rehmanniae radix, Paeoniae radix, Cnidii rhizoma, Angelicae radix, Menthae herba, Angelicae dahuricae radix, Saposhnikoviae radix, Forsythiae fructus, and Glycyrrhizae radix.
Amoxicillin and clavulanic acid used as reference antimicrobial drugs were purchased from Toronto Research Chemicals (Toronto, ON, Canada). Pure samples of apigenin, baicalein, baicalin, genistein, hesperidin, hesperetin, liquiritin, liquiritigenin, naringenin, glycyrrhizic acid, and glycyrrhetinic acid were purchased from Wako Pure Chemical Industries (Osaka, Japan). apigenin 7-O-glucoside was purchased from Sigma-Aldrich (St. Louis, MO, USA), narirutin from Extrasynthese (Genay, France), and isoliquiritin from Chemfaces (Hubei, China). baicalein, baicalin (baicalein 7-O-glucuronide), luteolin, genistin, wogonin, and wogonoside (wogonin 7-O-glucuronide), and berberine chloride were obtained from Tsumura & Co., with high purities for pharmacokinetic study.
Amoxicillin and clavulanic acid used as reference antimicrobial drugs were purchased from Toronto Research Chemicals (Toronto, ON, Canada). Pure samples of apigenin, baicalein, baicalin, genistein, hesperidin, hesperetin, liquiritin, liquiritigenin, naringenin, glycyrrhizic acid, and glycyrrhetinic acid were purchased from Wako Pure Chemical Industries (Osaka, Japan). apigenin 7-O-glucoside was purchased from Sigma-Aldrich (St. Louis, MO, USA), narirutin from Extrasynthese (Genay, France), and isoliquiritin from Chemfaces (Hubei, China). baicalein, baicalin (baicalein 7-O-glucuronide), luteolin, genistin, wogonin, and wogonoside (wogonin 7-O-glucuronide), and berberine chloride were obtained from Tsumura & Co., with high purities for pharmacokinetic study.
10
Modulating Wnt Signaling in HCEM Cells
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HCEM cells (1.0 × 105 cells per well) were seeded in 6-well plates (FALCON). After reaching confluence, FBS (Daiichi Chemical) levels were stepwise reduced in the medium to 0% as aforementioned, and then 100 ng/mL human recombinant Dickkopf (Dkk)-1, a Wnt signaling pathway antagonist (R&D system, Minneapolis, MN, USA) was added for 1 h before addition of 0–1 μM baicalin (Wako). Forty-eight hours after baicalin addition, we performed qPCR analysis as aforementioned.
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