Xanthohumol (Xn), purity >98%, was provided by the Chengdu Herbpurify CO., LTD (Chengdu, China). LPS (Escherichia coli 055:B5) and dimethyl sulfoxide (DMSO) were purchased from Sigma-Aldrich (St. Louis, MO, USA). Compound C and Brusatol (specific inhibitors of the AMPK, and Nrf2, respectively), and tert-butyl hydroperoxide (t-BHP) were offered by Sigma-Aldrich (St. Louis, MO). Penicillin and streptomycin, Fetal bovine serum (FBS) and Dulbecco's modified Eagle's medium (DMEM) were acquired from Invitrogen-Gibco (Grand Island, NY). Antibodies against Nrf2, Keap1, GCLC, HO-1, NQO1, GCLM, Trx-1, Txnip, NLRP3, casapase-1, ASC, IL-1β, Lamin B and β-actin were supplied by Cell Signaling (Boston, MA, USA) or Abcam (Cambridge, MA, USA). The Annexin V fluorescein isothiocyanate (FITC)/Propidium Iodide (PI) apoptosis kit were offered from Beyotime Institute of Biotechnology (Jinlin, China). In addition, GSH, MDA, SOD and MPO test kits were obtained from Nanjing Jiancheng Bioengineering Institute (Nanjing, China). All other chemicals were offered by Sigma-Aldrich (St. Louis, MO, USA), if not otherwise indicated.
Annexin 5 fluorescein isothiocyanate fitc propidium iodide apoptosis kit
Manufactured by Beyotime
Sourced in China
The Annexin V fluorescein isothiocyanate (FITC)/Propidium Iodide (PI) apoptosis kit is a laboratory reagent used for the detection and quantification of apoptosis in cells. It contains Annexin V-FITC, which binds to phosphatidylserine exposed on the cell surface during apoptosis, and Propidium Iodide, which stains the DNA of cells with compromised cell membranes.
Automatically generated - may contain errors
Lab products found in correlation
Penicillin and streptomycin, by Thermo Fisher Scientific (1 mentions)
Lamin b, by Cell Signaling Technology (1 mentions)
Casapase 1, by Cell Signaling Technology (1 mentions)
Mpo test kit, by Nanjing Jiancheng (1 mentions)
Txnip, by Cell Signaling Technology (1 mentions)
Lps escherichia coli 055 b5, by Merck Group (1 mentions)
Il 1β, by Cell Signaling Technology (1 mentions)
Dulbecco s modified eagle s medium dmem, by Thermo Fisher Scientific (1 mentions)
Fetal bovine serum (fbs), by Thermo Fisher Scientific (1 mentions)
β actin, by Cell Signaling Technology (1 mentions)
Tert butyl hydroperoxide tbhp, by Merck Group (1 mentions)
Dimethyl sulfoxide (dmso), by Merck Group (1 mentions)
Glutathione (gsh), by Nanjing Jiancheng (1 mentions)
Rpmi 1640 medium, by Sartorius (1 mentions)
Actin β polyclonal antibody, by Immunoway (1 mentions)
Fetal bovine serum (fbs), by Sartorius (1 mentions)
Facsverse, by BD (1 mentions)
4 protocols using annexin 5 fluorescein isothiocyanate fitc propidium iodide apoptosis kit
1
Xanthohumol Protects Against Oxidative Stress
2
Natural Compounds Modulate Apoptosis
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Quercetin, kaempferol, isorhamnetin, glycyrrhetinic acid, liquiritin, and glycyrrhizic acid ammonium salt were obtained from the National Institutes for Food and Drug Control (Beijing, China). Naringenin and licochalcone A were offered from Nanjing Spring and Autumn Biotech (Nanjing, China). Lipopolysaccharides (LPS; Escherichia coli 055: B5) were purchased from Solarbio LIFE SCIENCES (Beijing, China). The purity of each reference standard was above 98%. Cell Counting Kit 8 (CCK-8) was obtained from Dojindo Laboratory (Kumamoto, Japan). Fetal bovine serum (FBS) and RPMI 1640 medium were products of Biological Industries (Kibbutz Beit Haemek, Israel). Antibodies against Akt, Phospho-Akt (Ser473), Bcl-2, Bax and NF-κB p65 were supplied by Cell Signaling Technology (Beverly, MA, United States). Antibody for active + pro Caspase-3 was purchased from ABclonal Technology Co., Ltd. (Wuhan, China). Actin β Polyclonal Antibody was purchased from Immunoway (Suzhou, China). The Annexin V fluorescein isothiocyanate (FITC)/Propidium Iodide (PI) apoptosis kit was provided by the Beyotime Institute of Biotechnology (Shanghai, China). All other chemicals were provided by Sigma-Aldrich Co. (Shanghai, China), unless otherwise indicated.
3
Quantifying Apoptosis using Annexin V-FITC/PI Assay
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Annexin V-fluorescein isothiocyanate (FITC)-propidium iodide (PI) Apoptosis kit (Beyotime Institute of Biotechnology) was used to test cell apoptosis. Briefly, the EA.hy926 cells were seeded into 6-well plates at the density of 1 × 10 5 cells/well. After the cells well attached, 40 nM CdTe QDs was added to the wells and incubated for 24 hr. Then the cells were harvested and resuspended at a density 1 × 10 6 cells/mL in 500 μL binding buffer. According to the instruction of kit, 5 μL Annexin V-FITC was added to the samples and incubated at 4°C for 15 min. followed by staining with PI for 5 min. After that, the cells were resuspended in 500 μL PBS and analyzed by flow cytometry (BD-FACS-Verse). The fluorescence was monitored at 488 nm and 405 nm. Each plot represented 10,000 viable cells, and non-viable cells were excluded by appropriate gating. All data analyses were carried out using FCS express V3.0.
4
Cell Apoptosis Quantification by Flow Cytometry
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SRA01/04 cells were digested with trypsin after transfection, and the cell suspension was collected (1×10 5 cells). Then, Annexin V-fluorescein isothiocyanate (FITC)/propidium iodide (PI) Apoptosis Kit (Beyotime, Shanghai, China) was used to stain the cell suspension. Cell apoptosis rate was detected using a flow cytometer (BD Biosciences, San Jose, CA, USA).
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