The FITC-Annexin V apoptosis kit (KeyGen Biotechnology Co., Ltd.) was used to evaluate the apoptotic effects of Bs/CUS on HepG2 cells. Cells at a density of 4×104/ml per well were seeded in 6-well plates, incubated overnight at 37°C and treated for 72 h with Bs/CUS or kept untreated (UT) at concentrations of 1, 4, 16, and 64 nmol/l. Subsequently, the cells were collected, centrifuged at 500 × g for 5 min at room temperature, counted, 5 µl Annexin V-FITC and 5 µl PI were added for 15 min at room temperature in the dark for staining, resuspended in 500 µl binding buffer, and analyzed by flow cytometry (BD Biosciences).
Annexin 5 fitc apoptosis kit
Manufactured by Keygen Biotech
Sourced in China
The Annexin V-FITC Apoptosis Kit is a laboratory reagent used to detect and quantify apoptosis, a type of programmed cell death. The kit contains Annexin V conjugated with the fluorescent dye FITC, which binds to phosphatidylserine, a molecule that is exposed on the surface of apoptotic cells. This allows for the identification and enumeration of apoptotic cells using flow cytometry or fluorescence microscopy.
Automatically generated - may contain errors
Lab products found in correlation
Caspase 3 7 activity apoptosis assay kit, by AAT Bioquest (1 mentions)
Lipofectamine 2000, by Thermo Fisher Scientific (1 mentions)
Facscalibur flow cytometer, by Beckman Coulter (1 mentions)
Dual luciferase reporter assay kit, by Promega (1 mentions)
Caspase activity assay kit, by Beyotime (1 mentions)
Fetal bovine serum (fbs), by Thermo Fisher Scientific (1 mentions)
Oxaliplatin, by Merck Group (1 mentions)
Curcumin, by Merck Group (1 mentions)
Penicillin, by Beyotime (1 mentions)
Streptomycin, by Beyotime (1 mentions)
Cell counting kit 8 cck 8 kit, by Beyotime (1 mentions)
Calibur cytometer, by BD (1 mentions)
Trypsin without edta, by Beyotime (1 mentions)
Facscalibur system, by BD (1 mentions)
Facscalibur, by Bio-Rad (1 mentions)
Stattic, by Merck Group (1 mentions)
Gapdh, by Cell Signaling Technology (1 mentions)
Skov3 cells, by American Type Culture Collection (1 mentions)
Ros assay kit, by Beyotime (1 mentions)
Cellquest, by BD (1 mentions)
17 protocols using annexin 5 fitc apoptosis kit
1
Apoptotic Effects of Bs/CUS on HepG2 Cells
2
Apoptosis Evaluation of D-CUS245C on Lung Cancer Cells
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The FITC‐annexin V apoptosis kit (KGA106; KeyGEN BioTECH) was used to evaluate the apoptotic effects of D‐CUS245C on PD‐L1/SPC‐A‐1 and NC/SPC‐A‐1 cells. In preparation for the apoptosis assay, cells with a density of 5 × 104/mL per well were seeded in 6‐well plates for 24 hours. The cells were then treated with D‐CUS245C at concentrations of 0.5, 2.5, or 5 nmol/L for 72 hours, or kept untreated. After that, cells were collected, centrifuged, counted, resuspended in PBS, and analyzed with flow cytometry (BD Biosciences).
3
Assessing CD4+ T Cell Purity and Proliferation
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Flow cytometry was chosen to assess the purity of CD4+ T cells used for subsequent experiments. For the cell proliferation experiments, T cells were stained with CFSE before being seeded in 96-well plates and incubated for 96 h in the dark. T cells were collected and stained with Anti-CD4-APC antibody (dilution: 1:1000; Abcam) for 30 minutes. Cell proliferation was quantified by CFSE fluorescence using flow cytometry at 96 hours after treatment.
Similarly, apoptotic cells were analyzed using the FITC Annexin V Apoptosis Kit (KeyGEN BioTECH, China). The number of apoptotic cells, including both early and late apoptotic cells, was calculated.
Similarly, apoptotic cells were analyzed using the FITC Annexin V Apoptosis Kit (KeyGEN BioTECH, China). The number of apoptotic cells, including both early and late apoptotic cells, was calculated.
4
ER1626 Induces Apoptosis in Tumor Cells
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Tumor cells were grown to 80% confluency in 6-well plates, treated with ER1626 (10−7, 10−6 and 10−5 mol/L), and incubated for 24 h. Treated cells were processed with annexin V-FITC Apoptosis kit (KeyGEN, Nanjing, China) and were detected the distribution of cell state with FCM according to operation instruction.
5
Apoptosis Assay for Eca109 and KYSE30 Cells
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Apoptosis was assayed using the Annexin V-FITC Apoptosis Kit (Keygen, China) and Caspase 3/7 Activity Apoptosis Assay Kit (AAT Bioquest) according to the manufacturer's instructions. Briefly, the transfected Eca109 and KYSE30 cells were harvested and washed twice with PBS, followed by resuspension in Annexin-V-binding buffer. Then FITC-conjugated Annexin V and PI were added. After incubating for 10 min at room temperature in the dark, another binding buffer was added, and the samples were immediately analyzed using FACS Calibur flow cytometer (Beckman Coulter). For the caspase 3/7 activity apoptosis assay, cells were prepared and incubated with caspase 3/7 assay solution at room temperature for 1 h. Co-transfection of the reporter vector (pmirGLO-wt-SKP2 or pmirGLO-mut-SKP2) and miRNA (miR-186 mimics or scramble) was performed using Lipofectamine 2000 (Invitrogen). Twenty-four hours after transfection, firefly and Renilla luciferase activities were measured using the Dual Luciferase Reporter Assay Kit (Promega) according to the manufacturer's protocol.
6
Gastric Cancer Cell Line Assay Protocol
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Human gastric cancer cell line BGC-823 was obtained from cell resource center of Shanghai Biological Sciences Institute (Chinese Academy of Sciences, Shanghai, China).Cells were maintained in RMPI-1640 (Beyotime) supplemented with 10% fetal bovine serum (Invitrogen, Carlsbad, CA, USA), 100 U/ml penicillin (Beyotime), and 100 µg/ml streptomycin (Beyotime). The cultures were incubated at 37°C in a humidified atmosphere of 5% CO2.
Male BALB/c athymic nude mice (5 weeks old) were purchased from Shanghai Slac Laboratory Animal Co. Ltd (Shanghai, China). The protocol for the animal experiment was approved by the Institutional Animal Committee of Wenzhou University. All animals received care in accordance to the Guide for the Care and Use of Laboratory Animals [Permit No.: SYXK (zhe) 2010-0150].
5-FU, oxaliplatin, and curcumin were purchased from Sigma-Aldrich (St. Louis, MO, USA). Cell Counting Kit-8 (CCK-8 kit) and Caspase activity assay kits were purchased from Beyotime (Haimen, China). Annexin V-FITC Apoptosis Kit was purchased from Keygen Biotech (Nanjing, China).
Male BALB/c athymic nude mice (5 weeks old) were purchased from Shanghai Slac Laboratory Animal Co. Ltd (Shanghai, China). The protocol for the animal experiment was approved by the Institutional Animal Committee of Wenzhou University. All animals received care in accordance to the Guide for the Care and Use of Laboratory Animals [Permit No.: SYXK (zhe) 2010-0150].
5-FU, oxaliplatin, and curcumin were purchased from Sigma-Aldrich (St. Louis, MO, USA). Cell Counting Kit-8 (CCK-8 kit) and Caspase activity assay kits were purchased from Beyotime (Haimen, China). Annexin V-FITC Apoptosis Kit was purchased from Keygen Biotech (Nanjing, China).
7
Quantifying Cellular Apoptosis by FACS
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Percentages of apoptotic cells were evaluated by Annexin V-FITC apoptosis Kit (Keygen, China) according to manufacturer’s protocol. Flow cytometry was performed on a BD Calibur cytometer. Data were then obtained by FlowJo software.
8
Annexin V-FITC Apoptosis Assay
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An Annexin V-FITC Apoptosis Kit (KeyGen Biotech, Nanjing, China) was used to analyze cell apoptosis according to the manufacturer’s instructions. SMMC-7721 and HepG2 cells of logarithmic growth phase (1 × 105/ml) were seeded in six-well plates. Then, SMG9 NC or SMG9 siRNA was transfected into SMMC-7721 and HepG2 cells with Lipofectamine™ 3,000. Forty eight hours post-transfection, all cells (including cells in the supernatant) were harvested with trypsin without EDTA (Beyotime, Shanghai, China) and centrifuged at 1,200 rpm for 5 min. After washing with cold PBS twice, the cells were resuspended in 500 μl binding buffer and incubated with 5 μl Annexin V- FITC and 5 μl PI at room temperature for 15 min in the dark. Then, the samples were analyzed on a flow cytometer (BD FACS Calibur System; BD, United States).
9
Annexin V-FITC Apoptosis Assay
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We stained H9c2 cells with the Annexin V‐FITC Apoptosis Kit (KeyGEN, Jiangsu, China) following the protocols of manufacturer. The treated cells were rinsed using PBS, trypsinised with trypsin without EDTA, and subjected to centrifugation at 2000 revolutions per minute for 5 min at room temperature to harvest cells. We stained the obtained cells with 5 μL Annexin V-FITC and 5 μL of PI in a diluted solution and incubated them at room temperature in the dark for a quarter. Later, the stained cells were further detected and analysed using flow cytometry within half an hour (FACS Calibur, Bio-Rad Laboratories, Inc., USA). The lower left, upper left, lower right, and upper right quadrants, respectively, represent healthy, dead, early apoptotic, and late apoptotic cells.
10
Investigating STAT3 and NF-κB Signaling in SKOV3 Cells
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The antibodies for CD31, phospho-STAT3 (Tyr705), NF-kB (p65), GRP78, CHOP, and glyceraldehyde-3 phosphate dehydrogenase (GAPDH) were bought from Cell Signaling Technology. The antibodies for horseradish peroxidase-conjugated anti-mouse IgG and anti-rabbit IgG were bought from Epitomics. Stattic and PDTC were bought from Sigma. ELISA IL-6 kit and VEGF kit were purchased from R&D Systems. An Annexin V-FITC apoptosis kit was purchased from KeyGEN. SKOV3 cells from American Type Cell Culture (ATCC, Manassas, VA, USA) were grown in DMEM supplemented with 10% fetal bovine serum (FBS), 100 U/ml penicillin, 100 μg/ml streptomycin, and 25 μg/ml amphotericin B. Cells were cultured at 37°C in a humidified incubator with 5% CO2 and 95% air.
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