RPMI-1640 medium and fetal bovine serum (FBS) were purchased from Gibco (Grand Island, NY, USA). DAPI (4′,6-diamidino-2-phenylindole) was purchased from Wuhan Goodbio Technology Co., Ltd (Wuhan, China). The Annexin V-FITC Apoptosis Kit was purchased from BestBio (Shanghai, China). The Mitochondrial Membrane Potential Assay Kit was purchased from Signalway Antibody (College Park, MD, USA). The primary antibodies against poly(ADP-ribose) polymerase (PARP), caspase-3, cleaved caspase-3, PI3K-p110ɑ, p-AKT (Ser 473), AKT, mammalian target of rapamycin (mTOR), p-mTOR (S2448), p-p70S6, p70S6, p-S6 (240/244), p-S6 (235/236), S6, and Bcl-2 were purchased from Cell Signaling Technology (Beverly, MA, USA). The primary antibodies against X-linked inhibitor of apoptosis protein (XIAP), Bax, Noxa, Bcl-XL, Mcl-1, and β-actin were purchased from Abcam, Inc. (Cambridge, MA, USA). The human pancreatic cancer cell lines BxPC-3 (Catalog Number TCHu12) and SW1990 (Catalog Number TCHu201) were purchased from the Chinese Academy of Sciences (Shanghai, China). Cells were cultured in RPMI-1640 medium containing 10% FBS and 100 U/mL penicillin/streptomycin at 37 °C in 5% CO2 in a humidified atmosphere. Amcp was dissolved in dimethyl sulfoxide (DMSO) at a concentration of 10 mM. Gemcitabine was dissolved in DMSO at a concentration of 10 mM.
Annexin 5 fitc apoptosis kit
Manufactured by BestBio
Sourced in China, United States
The Annexin V-FITC Apoptosis Kit is a laboratory product designed to detect and quantify apoptosis, a programmed cell death process. It utilizes fluorescently labeled Annexin V, a protein that binds to phosphatidylserine, an important marker of apoptosis. The kit enables the identification and analysis of apoptotic cells through flow cytometry or fluorescence microscopy.
Automatically generated - may contain errors
Lab products found in correlation
Rpmi 1640 medium, by Thermo Fisher Scientific (6 mentions)
Mitochondrial membrane potential assay kit, by Signalway Antibody (4 mentions)
β actin, by Abcam (4 mentions)
Fetal bovine serum (fbs), by Thermo Fisher Scientific (4 mentions)
Mcl 1, by Abcam (3 mentions)
Pi3k p110, by Cell Signaling Technology (2 mentions)
Bcl xl, by Abcam (2 mentions)
Cleaved caspase 3, by Cell Signaling Technology (2 mentions)
Cycletest plus dna reagent kit, by BD (2 mentions)
Pvdf membrane, by Merck Group (2 mentions)
Bcl 2, by Abcam (2 mentions)
P akt ser473, by Cell Signaling Technology (1 mentions)
Bcl 2, by Cell Signaling Technology (1 mentions)
Tubulin polymerization assay kit, by Cytoskeleton (1 mentions)
Hoechst 33258, by Merck Group (1 mentions)
Beckman fc 500 flow cytometry, by Beckman Coulter (1 mentions)
Beckman fc500, by Beckman Coulter (1 mentions)
Cytomics fv 500, by Beckman Coulter (1 mentions)
Mtt powder, by Merck Group (1 mentions)
Pifithrin α, by Merck Group (1 mentions)
14 protocols using annexin 5 fitc apoptosis kit
1
Pancreatic Cancer Cell Apoptosis Assay
2
Gastric Carcinoma Cell Assays
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F12, RPMI-1640 medium and fetal bovine serum (FBS) were purchased from Gibco, BRL (Grand Island, NY, USA). The Cycletest Plus DNA Reagent Kit was purchased from BD Biosciences (Franklin Lakes, NY, USA). Hoechst33258 was obtained from Sigma-Aldrich (St Louis, MO, USA). The Annexin V-FITC Apoptosis Kit was purchased from BestBio (Shanghai, China). The Mitochondrial Membrane Potential Assay Kit was procured from Signalway Antibody (College Park, MD, USA). The Tubulin Polymerization Assay Kit was purchased from Cytoskeleton Inc (Denver, CO, USA). Primary antibodies were purchased from Abcam Inc (Cambridge, MA, USA). Human gastric carcinoma AGS and HGC-27 cell lines were purchased from the Chinese Academy of Sciences (Shanghai, China). Cells were cultured in F12 or RPMI-1640 medium containing 10% fetal bovine serum (FBS) and 1% penicillin/streptomycin at 37 °C in a 5% CO2 humidified atmosphere. Deacetylisovaltratum (DI) was dissolved in DMSO at a concentration of 100 mmol/L.
3
Quantifying Cell Apoptosis by Annexin V-FITC
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Cell apoptosis was quantified by using an Annexin V-FITC apoptosis kit (BestBio, China) according to the manufacturer’s instructions. NA cells were seeded into 6-well plates and incubated at 37°C overnight. Then cells were treated with RABV rHEP-Flury and P gene rearranged viruses at a MOI of 3. Twenty-four hours later, cells were collected and incubated with 5 μl Annexin V-FITC and 10 μl PI for 15 min. Finally, 500 μl of binding buffer was added to each tube and analyzed by a Beckman FC 500 flow cytometry(Beckman Coulter, Fullerton, CA, USA), followed by data analysis with the corresponding CXP Software.
4
RABV-Induced Apoptosis Evaluation
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NA cells were infected with RABV at an MOI of 3 and harvested at 24 hpi. Cells were stained with the Annexin V-FITC apoptosis kit (BestBio, Shanghai, China) according to the manufacturer’s protocols. Flow cytometry was performed on a Beckman FC500 flow cytometer (Beckman Coulter, Fullerton, CA, USA). Data were analyzed using CXP Software.
5
Quantifying Apoptosis via Annexin V
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Annexin V binding to HConF was performed using an Annexin V-FITC Apoptosis kit (BestBio) to measure apoptosis. HConFs (5×105/well) were plated into 6-well plates and then treated with medium containing different reagents for 48 h. Briefly, the cells, after being rinsed twice with phosphate-buffered saline (PBS), were resuspended in 400 µl of 1X binding buffer (10 mM HEPES, 140 mM NaCl, 2.5 mM CaCl2; pH 7.4), to which 5 µl of Annexin V-FITC was added and mixed well. After a 15-min incubation period at 2–8°C in the dark, 10 µl of propidium iodide (PI) was added to the cells and mixed well. The cells were incubated for a further 5 min at 2–8°C in the dark, after which time flow cytometry (Cytomics FV 500; Beckman Coulter, Brea, CA, USA) was performed within 15 min.
6
Curcumin Modulates Apoptosis Signaling
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Curcumin was purchased from Sigma-Aldrich (Shanghai, China). RPMI-1640 and penicillin/streptomycin were purchased from Invitrogen (Carlsbad, CA, USA), fetal bovine serum (FBS) was purchased from HyClone (UT, USA). Monoclonal antibodies specific for p38 mitogen-activated protein kinase (p38 MAPK); ERK1/2 and their phosphor-forms, and FOXO3a and p53 were purchased from Cell Signaling Technology (Beverly, MA, USA). The GAPDH, p53 and FOXO3a monoclonal antibodies were obtained from Abcam Co (Burlingame, CA, USA). PVDF membrane was purchased from Millipore (USA). PD98059 (a special inhibitor of ERK1/2) was purchased from Merck Millipore (Darmstadt, Germany), MTT powder and pifithrin-α (a special inhibitor of p53) were purchased from Sigma-Aldrich (St. Louis, MO, USA). p53 and FOXO3a siRNAs were obtained from Santa Cruz (CA, USA). Lipofectamine 2000 reagent was purchased from Invitrogen (Shanghai, China). The FOXO3a-GFP and N1-GFP plasmids were kindly provided by Frank M.J. Jacobs (Rudolf Magnus Institute of Neuroscience, Department of Pharmacology and Anatomy, University Medical Center, Utrecht) and was reported previously (14 (link)). Annexin V-FITC Apoptosis Kit was purchased from Bestbio Co. (Shanghai, China).
7
Cell Cycle and Apoptosis Analysis
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Cell cycle assays were performed using a Cell Cycle Analysis Kit (Beyotime). In brief, cells were collected, washed twice with PBS, and then fixed with pre-cooled 70% alcohol at 4 °C for 24 h. On the second day, the fixed cells were washed twice with cold PBS and then stained with PI, incubated at 37 °C in the dark for 30 min according to the manufacturer's instructions. Cell cycle distribution was determined using a FACS flow cytometer (BD Biosciences, San Jose, CA, US). The results were analyzed using Modifit software (version 5.0).
Apoptosis was detected using the Annexin V FITC Apoptosis Kit (BestBio, Shanghai, China). Cells were resuspended in 400 µL binding buffer, then 5 µL annexin V-FITC was added and incubated for 15 min at 4 °C in the dark, and then following the addition of 5 µL propidium iodide, were incubated for another 5 min in the dark; finally, the cells were analyzed by FACS flow cytometer. The analysis results were obtained using FlowJo 10.8.1 software.
Apoptosis was detected using the Annexin V FITC Apoptosis Kit (BestBio, Shanghai, China). Cells were resuspended in 400 µL binding buffer, then 5 µL annexin V-FITC was added and incubated for 15 min at 4 °C in the dark, and then following the addition of 5 µL propidium iodide, were incubated for another 5 min in the dark; finally, the cells were analyzed by FACS flow cytometer. The analysis results were obtained using FlowJo 10.8.1 software.
8
Apoptosis Signaling Pathway Assays
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RPMI‐1640 medium and foetal bovine serum (FBS) were purchased from Gibco (Grand Island, NY, USA). DAPI was purchased from Wuhan Goodbio Technology Co., Ltd (Wuhan, China). The Annexin V‐FITC Apoptosis Kit was purchased from BestBio (Shanghai, China). The Mitochondrial Membrane Potential Assay Kit was purchased from Signalway Antibody (College Park, MD, USA). The primary antibodies against poly(ADP‐ribose) polymerase (PARP), procaspase‐9, procaspase‐3, cleaved caspase‐3, PI3K‐p110ɑ, p‐mTOR (S2448), mTOR, p‐p70S6, p70S6, p‐S6 (240/244), p‐S6 (235/236), S6, p‐ERK and ERK were purchased from Cell Signaling Technology (Beverly, MA, USA). The primary antibodies against X‐linked inhibitor of apoptosis protein (XIAP), Bax, Noxa, Bcl‐XL, Bcl‐2, Mcl‐1 and β‐actin were purchased from Abcam Inc. (Cambridge, MA, USA).
9
Isoalantolactone's Effects on Pancreatic Cancer
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RPMI-1640 medium and fetal bovine serum (FBS) were purchased from Gibco (Grand Island, NY, USA). The Cycletest Plus DNA Reagent Kit was purchased from BD Biosciences (Franklin Lakes, NY, USA). DAPI was purchased from Wuhan Goodbio Technology CO., LTD (Wuhan, China). The Annexin V-FITC Apoptosis Kit was purchased from BestBio (Shanghai, China). Mitochondrial Membrane Potential Assay Kit was purchased from Signalway Antibody (College Park, MD, USA). The primary antibodies against Poly (ADP-ribose) polymerase (PARP), X-linked inhibitor of apoptosis protein (XIAP), Bax, Bak, Puma, Bcl-2, Mcl-1, AKT, p-AKT (S473, T308), mTOR, mTOR (S2448), p-Signal transducer and activator of transcription 3 (STAT3) and β-actin were purchased from Abcam Inc. (Cambridge, MA, USA).
Human pancreatic cancer cell line PANC-1 (Catalog No. SCSP-535) and SW1990 (Catalog No. TCHu201) cells were purchased from Chinese Academy of Sciences (Shanghai, China). Cells were cultured with DMEM or RPMI 1640 medium containing 10 % fetal bovine serum (FBS) and 1 % penicillin/streptomycin at 37℃, 5% CO 2 humidified atmosphere. Isoalantolactone (Catalog No. A1522, CAS No. 470-17-7) was purchased from Huzhou Zhanshu Biotechnology Co. Ltd (Zhejiang, China). F35
or isoalantolactone was dissolved in DMSO at the concentration of 100 mg/ml.
Human pancreatic cancer cell line PANC-1 (Catalog No. SCSP-535) and SW1990 (Catalog No. TCHu201) cells were purchased from Chinese Academy of Sciences (Shanghai, China). Cells were cultured with DMEM or RPMI 1640 medium containing 10 % fetal bovine serum (FBS) and 1 % penicillin/streptomycin at 37℃, 5% CO 2 humidified atmosphere. Isoalantolactone (Catalog No. A1522, CAS No. 470-17-7) was purchased from Huzhou Zhanshu Biotechnology Co. Ltd (Zhejiang, China). F35
or isoalantolactone was dissolved in DMSO at the concentration of 100 mg/ml.
10
Icaritin-Induced Cell Apoptosis
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Cells were seeded in six-well plates and incubated with various concentrations of Icaritin for 48 h. Cell apoptosis was determined using the annexin V-FITC apoptosis kit (BestBio, Shanghai, China) following manufacturer’s instructions. Cells were analyzed on a FACScan flow cytometer (BD Biosciences) and data were interpreted using the Flowjo software (Treestar).
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