CAPE (97%), CA (98%), FA (98%), EF (98%), caffeic acid methyl ester (CAME) (98%), caffeic acid ethyl ester (CAEE) (98%), calf thymus DNA (ct-DNA), ascorbic acid, and bovine serum albumin (BSA) were purchased from Sigma (St. Louis, MO, USA). Ferric chloride (≥99%), ferrous ammonium sulfate hexahydrate (≥99.5%), and trisodium citrate dihydrate (≥99.5%) were purchased from Sinopharm Chemical Reagent Co., Ltd. Calcein and Calcein-AM were purchased from MedChemExpress (MCE). 5,5-dimethyl-1-pyrroline N-oxide (DMPO) was purchased from DOJINDO Molecular Technologies.
Calcein am
Manufactured by MedChemExpress
Sourced in United States
Calcein-AM is a fluorescent dye used in cell viability and cytotoxicity assays. It is a non-fluorescent, cell-permeable compound that is converted to a green-fluorescent calcein upon hydrolysis by intracellular esterases in live cells. This conversion indicates the presence of metabolically active cells.
Automatically generated - may contain errors
Lab products found in correlation
Calcein, by MedChemExpress (1 mentions)
Ammonium ferrous sulfate hexahydrate, by Sinopharm (1 mentions)
Calf thymus dna ct dna, by Merck Group (1 mentions)
5 5 dimethyl 1 pyrroline n oxide, by Dojindo Laboratories (1 mentions)
Ascorbic acid, by Merck Group (1 mentions)
Bovine serum albumin (bsa), by Merck Group (1 mentions)
Ferric chloride, by Sinopharm (1 mentions)
Trisodium citrate dihydrate, by Sinopharm (1 mentions)
Deferiprone, by MedChemExpress (1 mentions)
Synergy 2, by Agilent Technologies (1 mentions)
Synergy ht, by Agilent Technologies (1 mentions)
Nh4cl, by Maclin Biochemical Technology (1 mentions)
Sorafenib, by MedChemExpress (1 mentions)
Wheat germ agglutinin alexa fluor 488 and 350 dyes, by Thermo Fisher Scientific (1 mentions)
Fecl3 6h2o, by Maclin Biochemical Technology (1 mentions)
Dcfh da, by Merck Group (1 mentions)
Sodium periodate, by Merck Group (1 mentions)
Dcfh da, by Solarbio (1 mentions)
α actinin, by Santa Cruz Biotechnology (1 mentions)
3 methyladenine 3 ma, by MedChemExpress (1 mentions)
8 protocols using calcein am
1
Antioxidant Assays with CAPE, Caffeic Acid, and DNA
2
Measuring Intracellular Lipid Levels
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The LIP levels were measured using calcein-acetoxymethyl ester (calcein-AM, MedChemExpress, NJ, USA) and deferiprone (MedChemExpress), according to the methods described in the literature [28 (link)]. BEAS-2B cells were seeded in 12-well plates at a density of 6 × 105 cells per well. After treated, cells were incubated with 0.05 μM Calcein AM for 15 min at 37°C. Then, cells were incubated with deferiprone for 1 h at 37°C. The medium containing deferiprone was replaced with fresh medium, and the cells were examined and imaged with a fluorescence microscope (BioTek Synergy 2).
3
Evaluating ABC Transporter Activity
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The activity of ABC transporters was evaluated by using Calcein-AM assay. For this assay, 6 × 104 per well THP-1 macrophages were seeded on 96-well black plates and then exposed to 0 (control), 6.25, 12.5, 25, 50, and 100 μg/mL Ag NMs with or without the presence of H3 for 24 hours. After exposure, the cells were thoroughly rinsed with Hanks solution, and then incubated with 1 μg/mL Calcein-AM (purchased from MedChemExpress, Monmouth Junction, New Jersey) in serum-free medium for 30 minutes. After that, the cells were rinsed again with Hanks solution, and the green fluorescence was read at Ex 485 ± 20 nm and Em 528 ± 20 nm by an ELISA reader (Synergy HT; BioTek, Tacoma, Washington).
4
Live/Dead Cell Imaging using Calcein-AM
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Calcein‐AM has very low toxicity, emits a strong green fluorescence signal after hydrolysis by lactonase, and can be used to observe the morphology of living cells. To prepare the live/dead working solution, 4 μl of ethidium homodimer‐1 and 1 μl of Calcein‐AM (MedChemExpress, China, Cat.HY‐D0041) were mixed with 2 ml of phosphate‐buffered saline (PBS). Then, 0.1 ml of live/dead working solution was added to the 96‐well plate created in the previous section and incubated at 37°C in the dark for 30 min. After two PBS washes, samples were analyzed by confocal microscopy (100x, Leica; Deerfield, IL, USA) equipped with a fluorescence light source. A commercial polystyrene‐coated cell culture plate was used as a control.
5
Sorafenib-Induced Oxidative Stress Assay
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Sorafenib, Calcein-AM and TMRE were purchased from Med Chem Express (MCE). DCFH-DA (2'-7'-dichlorodihydrofluorescein diacetate) was purchased from Sigma. FeCl3· 6H2O and NH4Cl were purchased from MACKLIN. Sulphosalicylic acid was purchased from Aladdin. Wheat germ agglutinin (WGA) Alexa Fluor 488 and 350 dyes were purchased from Thermo Scientific. GFP and OFP fluorescin antibodies for Western blotting were purchased from Transgen. COX2 antibodies for Western blotting and TFRC, Ki67 antibodies for IHC were purchased from Cell Signaling Technology.
6
Multifunctional Hybrid Hydrogel for Tissue Regeneration
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Dextran (Dex, Mw = 200 kDa, Psaitong, China), hyaluronic acid sodium salt (HA, Mw = 50 kDa, Heowns, China), sodium periodate (99%, Sigma-Aldrich, USA), carbohydrazide (CDH, 97%, Heowns, China), resveratrol (Res, 98%, Heowns, China), 2-hydroxypropyl-β-cyclodextrin (HP-β-CD, 98%, Heowns, China), ethylene glycol (99%, Aladdin, China), 1-hydroxybenzotriazole (HOBT, 98%, Heowns, China), N-(3-dimethylaminopropyl)-N′-ethylcarbodiimide hydrochloride (EDC, aladdin, China), hydrogen peroxide (H2O2, 30 wt%, aladdin, China), 3-methyladenine (3-MA, 99%, MedChemExpress, USA), propidium iodide (PI, 99.44%, MedChemExpress, USA), Calcein acetoxymethyl ester (Calcein-AM, 95%, MedChemExpress, USA), 2′,7′-dichlorodihydrofluorescein diacetate (DCFH-DA, Solarbio, China), 5,5′,6,6′-tetrachloro-1,1′,3,3′-tetraethyl-imidacarbocyanine (JC-1, 99%, MedChemExpress, USA), mitochondrial superoxide indicator (MitoSOX Red, Yeasen, China), 1-diphenyl-2-picrylhydrazyl free radical (DPPH, 97%, TCI, China) and 3-(4,5-dimethyl-2-thiazoyl)-2,5-diphenyl tetrazolium bromide (MTT, 98%, Solarbio, China) were used as received. Antibodies of α-actinin, vWF and α-SMA were purchased from Santa Cruz Biotechnology (USA). The antibodies used in western blot were all obtained from Abcam (UK). All the other reagents were of analytical grade and used without further purification.
7
Quantifying Tube Formation of HLECs in Matrigel
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HLECs (8 × 104) suspended in CM from ccRCC cells were seeded into a Matrigel‐coated 48‐well plate. After 8 h culture, HLECs were washed with phosphate‐buffered saline (PBS) and stained with Calcein‐AM (#HY‐D0041, MedChemExpress). Then the tube‐like structure on the growth factor‐reduced Matrigel was photographed by a microscope (#DMI3000B, Leica, Wetzlar, Hessen, Germany). The total tube length and the number of junctions were analyzed using the ImageJ software (National Institutes of Health, Bethesda, MD, USA).
8
Cellular Migration Driven by Fluid Shear
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NIH-3T3 cells were seeded at a density of 5 × 105/ml onto the plane or groove substrates in 24-well plates and grown to 100% confluence after 8 h of culture. Then, the samples were cultured in serum-free medium. The cell monolayer was scratched with a sterile 200-μl pipette tip and washed with PBS to remove cellular debris. For groove specimens, the scratch direction is parallel or vertical to the groove structure. After 24 h of working with the 0.6-Hz fluid shear force parallel or perpendicular to the scratch, a new scratch was made parallel to the direction of the first one on each substrate as the initial width, and then stained with Calcein-AM (MedChemExpress, Monmouth Junction, USA) with a final concentration of 0.57μM. The changes in cellular migration were recorded with a fluorescence microscope (Carl Zeiss, Oberkochen, Germany) at low magnification (5×). For each image, distances between one side of scratch and the other were quantified using the NIH ImageJ software, and the spacing was normalized to initial width. The assay was repeated four times.
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