Emodin and aloe-Emodin were purchased from Sigma-Aldrich (MO, USA) and dissolved in DMSO. The studied concentrations were chosen according to the cytotoxicity assay which was done in the range of 0-1000 µM with the ATP assay. According to the IC50 values, for the evaluation of the gene expression levels 25 µM, 12.5 µM, and 6.25 µM were selected for Emodin; 10 µM, 5 µM, and 2.5 µM for aloe-Emodin. The control group was the solvent containing the same percentage of DMSO (0.1%) with the treatment groups.
Aloe emodin
Manufactured by Merck Group
Sourced in United States
Aloe-emodin is a chemical compound found in the aloe vera plant. It is a naturally occurring anthraquinone derivative. Aloe-emodin can be used as a reference standard or in research applications involving the analysis of aloe vera extracts or related compounds.
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Lab products found in correlation
Emodin, by Merck Group (6 mentions)
Rhein, by Merck Group (4 mentions)
Chrysophanol, by Merck Group (4 mentions)
Methanol, by Merck Group (3 mentions)
Milli q water purification system, by Merck Group (3 mentions)
Physcion, by Merck Group (2 mentions)
Sennoside a, by Merck Group (2 mentions)
Thioflavin t, by Merck Group (2 mentions)
Formic acid, by Merck Group (2 mentions)
Acetonitrile, by Merck Group (2 mentions)
Fdr 50p freeze dryer, by Iwaki (1 mentions)
Whatman, by Cytiva (1 mentions)
Rt pcr kit, by Promega (1 mentions)
Trizol, by Thermo Fisher Scientific (1 mentions)
Acetonitrile, by Avantor (1 mentions)
Isoliquiritigenin, by Merck Group (1 mentions)
Liquiritigenin, by Merck Group (1 mentions)
Anti claudin 5, by Santa Cruz Biotechnology (1 mentions)
Methanol, by Junsei (1 mentions)
Nunc plate, by Thermo Fisher Scientific (1 mentions)
14 protocols using aloe emodin
1
Cytotoxic Evaluation of Emodin and Aloe-emodin
2
HPLC-Grade Solvent Purification
3
Quantification of Anthraquinones in R. palmatum
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Crude R. palmatum extract powder was obtained from Sun Ten Pharmaceutical Co., Ltd. For each tested extract, 1 g of powder was dissolved in 40 ml methanol or distilled water and gently shaken overnight at room temperature. Solution samples were filtered with Whatman No. 1 filter paper, lyophilized in an IWAKI FDR-50P freeze dryer, then sterilized by a 0.44 μm syringe filter and stored at −80 °C until used (100 % stock solution). Also, filtered extract was injected directly into the HPLC instrument with C-18 reverse phase column. Separation was conducted with gradient elution using acetonitrile and 0.1 % phosphoric acid at a flow rate of 1 ml/min, eluent detected at 250 nm. Chrysophanol, rhein, emodin, aloe-emodin, and physcion were purchased from Sigma Chemical Co. of St. Louis, serving as external standards for comparing chromatographic peaks of methanol and water extracts with the retention time of these marker compounds.
4
Analytical Profiling of Herbal Compounds
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Anti-claudin 5 was obtained from Santa Cruz Biotechnology (Santa Cruz, CA, USA). Horseradish peroxidase conjugated goat anti-rabbit and goat anti-mouse IgGs were purchased from Novus Biological (Centennial, CO, USA). Trizol was obtained from Invitrogen (Invitrogen, Carlsbad, CA, USA) and reverse transcriptional polymerase chain reaction (RT-PCR) kit was obtained from Promega (Promega, Madison WI, USA). Sennoside A, emodin, chrysophanol, aloe-emodin, rhein, glycyrrhizin acid, liquiritigenin, isoliquiritigenin, and other reagents were purchased from Sigma-Aldrich (St. Louis, MO, USA). The reagents for ultra-performance liquid chromatography (UPLC) analysis were methanol (Junsei for the high-performance liquid chromatography (HPLC), acetonitrile (JT BAKER for the HPLC), and then Water (Tertiary distilled water).
5
Aloe-emodin Cytotoxicity in HeLa Cells
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The HeLa cell line (human cervix carcinoma) was cultured in Nunc plates at a temperature of 37 °C and in a 5% carbon dioxide atmosphere in a CO2 DirectHeat incubator (Thermo Fisher Scientific). Cells came from the Department of Radiobiology and Immunology, UJK Kielce. Cell culture was carried out in DMEM medium supplemented with 10% fetal bovine serum (FBS) and 1% antibiotic mixture from Thermo Fisher. Aloe-emodin (C15H10O5) was purchased from Sigma-Aldrich (USA). Cells were exposed to the test anthraquinone in concentration ranges of 1 μM to 100 μM.
6
Screening G-Quadruplex Ligand Compounds
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Pyridostatin (PDS) (33 (link)), hemin (34 (link)), bisquinolinium derivative PhenDC3 (35 (link)), thioflavin T (ThT) (36 (link)), thiazole orange (TO) (37 (link)), crystal violet (CV) (38 (link)), acridine derivative BRACO-19 (39 (link)), hydroxyantraquinone natural compound aloe-emodin (40 (link)) and natural alkaloid berberine (41 (link)) were purchased from Sigma-Aldrich (Prague, Czech Republic). N-Methylmesoporphyrin IX (NMM) (42 (link),43 ) and 5,10,15,20-tetrakis-(N-methyl-4-pyridyl)porphine (TMPyP4) (42 (link),44 (link)) were obtained from Santa Cruz Biotechnology (Heidelberg, Germany). Carboxy Pyridostatin (cPDS) (45 (link),46 (link)), bisquinolinium derivative 360A (47 (link)) and polymerase I inhibitor CX5461 (48 (link)) were from MedChemExpress (Stockholm, Sweden). Test compounds were solubilized according to the recommendations of the provider in water or in 100% (v/v) dimethyl sulfoxide (DMSO) to yield stock solutions of desired concentration, usually 10 mM.
7
Phytochemical Compounds from Natural Sources
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Silybin, 7-hydroxyflavone, flavanone, saponin, lupeol, gluconic acid, galacturonic acid, D-sorbitol, digitonin, arbutin, D- (-) salicin, kaempferitrin, isoquercitrin, chrysophanic acid, aloe-emodin, o-coumaric acid, and vanillin were purchased from Sigma, St. Louis, MO, USA. Pinocembrin, β-sitosterol, and β-sitosterol-O-glucoside were isolated from Centaurea eryngioides [25 ]. Glucuronic acid and ouabain were obtained from Serva, Feinbiochemica, Heidelberg, Germany. Naringin was isolated from the peel of Citrus jambhiri Lush. fruit [26 (link)]. The investigated phytochemicals in the current study are discussed in Table 1 .
8
Analytical Method for Aloe and Senna Compounds
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Aloin A, aloin B, aloe emodin, emodin, and danthron were purchased from Sigma-Aldrich (Milan, Italy). LC-grade acetonitrile, formic acid, and methanol were obtained from Merck KGaA (Darmstadt, Germany). Ultrapure water (18.2 MΩ·cm) was produced using a Milli-Q water purification system from Millipore (Milford, MA, USA). Dry Cassia senna L. leaf and fruit extracts (dry extracts A–C), tablets containing extracts of C. senna L. and Aloe, and liquid products (Aloe juices A–C and hydroalcoholic solutions in the form of oral spray and single-dose bottles) with Aloe and C. senna L. were obtained from external companies. The latter were used to assess the robustness (i.e., the capacity to remain unaffected by small, deliberate variations in method parameters) and sensitivity of the analytical method.
9
HPLC Solvent Preparation and Compound Procurement
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Water for high-performance liquid chromatography (HPLC) was purified by reverse-osmosis. All other solvents were HPLC grade and were purchased from Fisher Scientific. Aloe-emodin was purchased from Sigma-Aldrich.
10
Aloe-emodin Bioactivity in Cell Lines
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Aloe-emodin was originally isolated from leaves of Aloe vera (Hamman, 2008 (link)). In the present study, we purchased Aloe-emodin from Sigma Chemical Co., St. Louis, MO, USA. All other chemicals used in this study were of the molecular biology grades and commercially available.
RIN5F and L6 myotubes were obtained from American Type culture collections (ATCC, Manassas, VA 20108 USA). Cell culture materials such as, RPMI-1640 (AG Biochrom, Germany), Fetal bovine serum (FBS) (Hiclone, Germany) and streptomycin (AG Biochrom, Germany. All spectrophotometric measurements were carried out using UV2010 Spectrophotometer (Hitachi, Germany). The cell line was maintained and the experiments were carried out according to the guidelines of CMRC ethical committee of KKUH, Saudi Arabia.
RIN5F and L6 myotubes were obtained from American Type culture collections (ATCC, Manassas, VA 20108 USA). Cell culture materials such as, RPMI-1640 (AG Biochrom, Germany), Fetal bovine serum (FBS) (Hiclone, Germany) and streptomycin (AG Biochrom, Germany. All spectrophotometric measurements were carried out using UV2010 Spectrophotometer (Hitachi, Germany). The cell line was maintained and the experiments were carried out according to the guidelines of CMRC ethical committee of KKUH, Saudi Arabia.
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