For the cell cycle analyses, the ZLD1039-treated cells were fixed and stained with the PI staining solution for 10 min in the dark and then analysed using flow cytometry (FCM). The data were analysed using the Cell Quest and Flow Jo software. For the apoptosis assays, the cells were harvested and analysed using an Annexin V- fluorescein isothiocyanate (FITC) apoptosis detection kit (Roche, Indianapolis, IN, USA) according to the manufacturer’s protocol.
Annexin 5 fluorescein isothiocyanate fitc apoptosis detection kit
Manufactured by Roche
Sourced in United States, Germany
The Annexin V-fluorescein isothiocyanate (FITC) apoptosis detection kit is a laboratory reagent used to detect and quantify apoptosis, a programmed cell death process, in cell samples. It contains Annexin V, a protein that binds to phosphatidylserine, a marker of apoptotic cells, and is conjugated to the fluorescent dye FITC for visualization and analysis using techniques such as flow cytometry or fluorescence microscopy.
Automatically generated - may contain errors
Lab products found in correlation
Cell counting kit 8 cck 8, by Beyotime (1 mentions)
2 7 dichlorofluorescein diacetate dcfh da, by Beyotime (1 mentions)
Ab13847, by Abcam (1 mentions)
Ab192591, by Abcam (1 mentions)
Ab21685, by Abcam (1 mentions)
Ab48187, by Abcam (1 mentions)
Ab37149, by Abcam (1 mentions)
Ab124956, by Abcam (1 mentions)
Ab11419, by Abcam (1 mentions)
Ab37073, by Abcam (1 mentions)
Ab179461, by Abcam (1 mentions)
Ab79483, by Abcam (1 mentions)
Facscanto ll, by BD (1 mentions)
4 protocols using annexin 5 fluorescein isothiocyanate fitc apoptosis detection kit
1
Cell Cycle and Apoptosis Analysis
2
Endoplasmic Reticulum Stress Pathway
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Antibodies against phosphorylated (p)-protein kinase R-like ER kinase (PERK; ab192591), PERK (ab79483), inositol-requiring kinase 1α (IRE1α; ab37073), p-IRE1α (ab48187), activating transcription factor 6α (ATF6α), p-50ATF6α (ab37149), glucose-regulated protein 78 (GRP78; ab21685), CCAAT/enhancer binding protein homologous protein (CHOP; ab11419), p-c-Jun N-terminal kinase (JNK; ab124956), JNK (ab179461) and caspase-3 (ab13847) were purchased from Abcam (Cambridge, UK). N-acetyl L-cysteine (NAC), 4-phenylbutyric acid (4-PBA) and thap-sigargin (Thap) were also purchased from Abcam. The Annexin V-fluorescein isothiocyanate (FITC) apoptosis detection kit was purchased from Roche Diagnostics (Indianapolis, IN, USA). Cell Counting kit-8 (CCK-8) and 2′,7′-dichlorofluorescein diacetate (DCFH-DA) were purchased from Beyotime Institute of Biotechnology (Shanghai, China). SW (osmolality, 1,300 mmol/l; pH 8.2; relative density 1.05; salt content, 34.421%; NaCl, 26.518 g/l; MgSO4, 3.305 g/l; MgCl2, 2.447 g/l; CaCl2, 1.141 g/l; KCl, 0.725 g/l; NaHCO3, 0.202 g/l; and NaBr, 0.083 g/l) was prepared based on the overall composition of the East China Sea, which was provided by the Chinese Ocean Bureau (Beijing, China).
3
Annexin V-FITC Apoptosis Assay
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Cell apoptosis was assessed using an annexinV-fluorescein isothiocyanate (FITC) apoptosis detection kit (Roche Diagnostics, Germany) according to the manufacturer’s instructions. Briefly, after the treatment, astrocytes were washed with PBS and collected into new tubes. After gently resuspending in annexinV binding buffer, the cells were incubated with annexinV-FITC/PI in the dark for 15 min and detected by flow cytometry using a BD FACSCantoll (Becton, Dickinson and Company, USA).
4
Quantification of Ethanol-Induced Apoptosis
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To further examined whether MFA could L-02 cell apoptosis induced by ethanol, apoptosis was quantified using an Annexin V-fluorescein isothiocyanate (FITC) Apoptosis Detection kit (Roche Diagnostics GmbH, Mannheim, Germany) in accordance with the manufacturer's instructions. Briefly, following treatment with MFA for 24 h at various concentrations as mentioned above, the cells were harvested and washed twice with cold PBS. Following centrifugation (500 x g, 5 min, 25˚C), the cells were stained with Annexin V-FITC and PI, and then analyzed by flow cytometry (flow cytometer; BD Biosciences, San Jose, CA, USA).
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