The MDA-MB-231 and SKBR3 human breast cancer cell lines were cultured in Dulbecco's modified Eagle's medium (DMEM, Gibco; Thermo Fisher Scientific, Inc., Waltham, MA, USA) supplemented with 10% fetal bovine serum (FBS; Invitrogen; Thermo Fisher Scientific, Inc.), penicillin (100 U/ml) and streptomycin (100 U/ml) in a humidified atmosphere containing 5% CO2 at 37°C. Genistein, calcein-acetoxymethyl (AM) and MTT were purchased from Sigma-Aldrich (Merck KGaA, Darmstadt, Germany). An Annexin V-FITC/PI Apoptosis Detection Kit, Transwell inserts and Matrigel were purchased from BD Biosciences (Franklin Lakes, NJ, USA). Primary antibody against tubulin (cat no. SC-5274) was purchased from Santa Cruz Biotechnology, Inc. (Dallas, TX, USA). Anti-p21 (cat no. 2946), anti-Skp2 (cat no. 4358), and anti-p27 (cat no. 2552) antibodies were obtained from Cell Signaling Technology, Inc. (Danvers, MA, USA). Anti-mouse HRP-linked antibody (cat no. 7076) and anti-rabbit HRP-linked antibody (cat no. 7074) were purchased from Cell Signaling Technology, Inc.
Calcein acetoxymethyl am
Manufactured by Merck Group
Sourced in United States
Calcein-acetoxymethyl (AM) is a cell-permeant, fluorescent dye used for the detection and quantification of intracellular calcium levels. It serves as a calcium indicator, allowing for the monitoring of calcium dynamics within living cells.
Automatically generated - may contain errors
Lab products found in correlation
Genistein, by Merck Group (1 mentions)
Anti skp2, by Cell Signaling Technology (1 mentions)
Anti rabbit hrp linked antibody, by Cell Signaling Technology (1 mentions)
Anti p27, by Cell Signaling Technology (1 mentions)
Fetal bovine serum (fbs), by Thermo Fisher Scientific (1 mentions)
Methyl thiazolyl tetrazolium (mtt), by Merck Group (1 mentions)
Matrigel, by BD (1 mentions)
Annexin 5 fitc pi apoptosis detection kit, by BD (1 mentions)
Transwell insert, by BD (1 mentions)
Phosphate buffer saline pbs ph 7.4, by Solarbio (1 mentions)
Tcs sp8, by Leica (1 mentions)
Penicillin streptomycin, by Solarbio (1 mentions)
Hydrochloric acid (hcl), by Sinopharm (1 mentions)
Urea, by Sinopharm (1 mentions)
Citric acid, by Sinopharm (1 mentions)
3 protocols using calcein acetoxymethyl am
1
Genistein Induces Apoptosis in Breast Cancer Cells
2
Cytotoxicity Evaluation of Chemical Reagents
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Urea, citric acid, and hydrochloric acid were purchased from Sinopharm Group Chemical Reagent (Shanghai, China). Cell counting kit 8 (CCK-8), Hoechst 33342, propidium iodide (PI), trypsin, penicillin/streptomycin, and phosphate buffer saline (PBS; pH 7.4) were obtained from Solarbio (Beijing, China). Calcein acetoxymethyl (AM) was obtained from Sigma-Aldrich (St. Louis, MO, USA). Dulbecco’s modified Eagle’s medium (DMEM) and RPMI-1640 medium were purchased from Yuanpei Biotechnology Co., Ltd. (Shanghai, China). Fetal bovine serum (FBS) was purchased from PAN Seratech (Leica TCS SP8, Wetzlar, Germany).
3
Visualizing Cell Cultures with Calcein-AM
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Cells cultured inside the device were stained with Calcein-acetoxymethyl (AM) (Sigma Aldrich, St. Louis, MO, USA) for visualization and imaged with a fluorescent microscope (IX71, Olympus, Center Valley, PA, USA) equipped with a camera (DP70, Olympus). First, cells were briefly rinsed with a phosphate buffered saline (PBS) and 1 μM of Calcein-AM was added to both the soma compartment and the axon compartments followed by incubation in a 37 °C humidified incubator for 10 min. Cells were rinsed with PBS three more times before acquiring images.
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