DOX hydrochloride was purchased from Nanjing Oddfoni Biological Technology Co., Ltd (Nanjing, China). The siRNAIGF1R with the sequence of 5′-CAUACUGCGCUCUAUAGAUTT-3′ was selected for targeting the IGF1R gene. The double-stranded siRNAIGF1R and FAM-labeled siRNAIGF1R (FAM: carboxyfluorescein) were designed and chemically synthesized by GenePharm Co., Ltd (Shanghai, China). 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) cell proliferation and cytotoxicity assay kit, Annexin V-FITC apoptosis detection kit, Hoechst 33342, 4% paraformaldehyde, folate, biotin, RPMI-1640 medium, phosphate-buffered saline (PBS), fetal bovine serum (FBS), bicinchoninic acid (BCA) protein assay kit and polyvinylidene difluoride (PVDF) membrane were provided by Sangon Biotech Co., Ltd (Shanghai, China). Mouse monoclonal IGF1R beta chain antibody and HRP-conjugated beta-actin antibody, HRP-conjugated goat anti-mouse IgG (H + L) were from Proteintech Group, Inc (USA). All other reagents used were analytical grade and ultrapure water (18.25 MΩ) was used throughout the study.
Annexin 5 fitc apoptosis detection kit
Manufactured by Sangon
Sourced in China
The Annexin V-FITC Apoptosis Detection Kit is a laboratory tool used to detect and quantify apoptosis, a type of programmed cell death. The kit utilizes Annexin V, a protein that binds to phosphatidylserine, which is exposed on the surface of apoptotic cells. Annexin V is conjugated with the fluorescent dye FITC, allowing for the visualization and measurement of apoptotic cells using flow cytometry or fluorescence microscopy.
Automatically generated - may contain errors
Lab products found in correlation
Paraformaldehyde (pfa), by Sangon (1 mentions)
Rpmi 1640 medium, by Sangon (1 mentions)
Hoechst 33342, by Sangon (1 mentions)
Facs flow cytometry, by BD (1 mentions)
Calcein am, by Thermo Fisher Scientific (1 mentions)
Bovine serum albumin (bsa), by Merck Group (1 mentions)
Sybr gold, by Thermo Fisher Scientific (1 mentions)
Propidium iodide, by Merck Group (1 mentions)
Hoechest 33342, by Thermo Fisher Scientific (1 mentions)
Dulbecco s phosphate buffered saline dpbs, by Corning (1 mentions)
Caspase 3 detection kit, by Beyotime (1 mentions)
Doxorubicin dox, by Dingguo (1 mentions)
Lsrii flow cytometer system, by BD (1 mentions)
2 iminothiolane hydrochloride traut s reagent, by Merck Group (1 mentions)
Fetal bovine serum (fbs), by Wisent (1 mentions)
Anti cd8 antibody, by Abcam (1 mentions)
Anti cd4 antibody, by Abcam (1 mentions)
Accuri c6 flow cytometer, by BD (1 mentions)
Cellquest pro software, by BD (1 mentions)
Cytoflex flow cytometer, by Beckman Coulter (1 mentions)
12 protocols using annexin 5 fitc apoptosis detection kit
1
Targeting IGF1R with siRNA in Cancer Cells
2
Investigating Sorafenib and Metformin Effects on MHCC97H Cell Apoptosis
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MHCC97H cells were plated in 96-well plates at 4000 cells/well (n = 6) containing 100 μl of DMEM +10 % FBS treated with 400 μM CoCl2 and cultured for 24 h, then incubated with sorafenib or metformin for another 48 h. DMEM (100 μl) and CCK8 (10 μl) were added to each well and incubated for 2 h. Then, the absorbance was detected with a microplate reader at a test wavelength of 450 nm.
Annexin V/PI was applied to investigate the impact of sorafenib or metformin on cell apoptosis. After treatments, the cells were added with 5 μl Annexin V and 10 μl PI staining supplied by the Annexin V-FITC Apoptosis Detection Kit (SANGON, Cat. No. BS6336), then the results were measured by flow cytometry using a FACS flow cytometer (Becton Dickinson verse, San Jose, CA).
Annexin V/PI was applied to investigate the impact of sorafenib or metformin on cell apoptosis. After treatments, the cells were added with 5 μl Annexin V and 10 μl PI staining supplied by the Annexin V-FITC Apoptosis Detection Kit (SANGON, Cat. No. BS6336), then the results were measured by flow cytometry using a FACS flow cytometer (Becton Dickinson verse, San Jose, CA).
3
Evaluating Cell Viability and Apoptosis
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Microplates containing 96 wells were loaded with 4000 MHCC97H cells/well (n=6) along with DMEM (10 ml), FBS (10%), and 400 mM CoCl2 solution and subjected to incubation for 24 h. After 24 h, the cells were treated with RGF and MTF and incubated for 48 h. Later, in each well, 100 ml DMEM and 10 ml CCK8 solutions were added, and after 2 h the absorbance in each plate was measured at λmax450 nm.
To study the effect of RGF and MTF on cell apoptosis, Annexin V and PI (10ml) staining (bought from Annexin V-FITC Apoptosis detection kit, SANGON, Cat. No. BS6336) was applied to the cells and the effects were analyzed using FACS flow cytometry (Becton Dickinson, San Jose, CA).
To study the effect of RGF and MTF on cell apoptosis, Annexin V and PI (10ml) staining (bought from Annexin V-FITC Apoptosis detection kit, SANGON, Cat. No. BS6336) was applied to the cells and the effects were analyzed using FACS flow cytometry (Becton Dickinson, San Jose, CA).
4
Apoptosis Assay of NSCLC Cells
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Following transfection for 48 h, an apoptosis assay of A549 and H1975 cells (1×106 cells) was performed using an Annexin V-FITC Apoptosis Detection kit (Sangon Biotech Co., Ltd.). A FACSCant II flow cytometry system (BD Biosciences) was used to monitor apoptosis (FlowJo v10.0.7; FlowJo LLC). Early apoptotic NSCLC cells were Annexin V (+)/propidium iodide (PI) (−) and presented in the lower right quadrant, whereas late apoptotic NSCLC cells were Annexin V (+)/PI (+) and presented in the upper right quadrant.
5
Annexin V-FITC Apoptosis Assay
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The cells for the apoptosis analysis were stained with anti-Annexin V antibody and propidium iodide using an Annexin V-FITC Apoptosis Detection kit (Sangon Biotech, Shanghai, China), and the percentage of apoptotic cells was examined with flow cytometry (BD Bioscience, CA, USA).
6
Cell Viability and Apoptosis Assay
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All DNA were synthesized and purified by Sangon Biotechnology Company, Ltd. (Shanghai). Sequences of all the DNA were listed in Supplementary Table S1 . Dulbecco’s phosphate buffered saline (D-PBS) was purchased from Corning. 0.1 M PB buffer was prepared using NaH2PO4 and Na2HPO4. Binding buffer was D-PBS containing 5 mM MgCl2, 4.5 g/L glucose, 1 mg/ml BSA and 0.1 mg/ml yeast tRNA. Methylthiazolyldiphenyl-tetrazolium bromide (MTT), yeast tRNA, bovine serum albumin (BSA) and propidium iodide (PI) were purchased from Sigma-Aldrich. Calcein-AM, SYBR Gold and Hoechest 33,342 were purchased from Thermo Fisher Scientific. Doxorubicin (Dox) was purchased from Dingguo Biotechnology Company, Ltd. (Beijing). Caspase 3 detection kit was purchased from Beyotime Biotechnology Company, Ltd. (Shanghai). Annexin-V FITC apoptosis detection kit was purchased from Sangon Biotechnology Company, Ltd. (Shanghai). Other chemicals were of analytical grade and used without further purification.
7
Quantifying Cell Apoptosis with Flow Cytometry
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Cell apoptotic rate was determined using flow cytometry via an Annexin V-FITC Apoptosis Detection Kit (Sangon Biotech, Shanghai, China). Generally, cells were re-suspended in 195 µL of binding buffer (1×) at a density of 2−5×105 cells/mL. Then, 5 µL of Annexin V-FITC solution was added to each sample for 15 min at room temperature in the dark. After washing with binding buffer (1×), 10 µL of propidium iodide solution was added to cell samples [re-suspended in 195 µL of binding buffer (1×)]. Finally, cell apoptotic rate was determined by flow cytometry (BD Biosciences, San Jose, CA, USA).
8
Apoptosis Detection by Annexin V-FITC
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Cell apoptosis was evaluated by using the Annexin V-FITC Apoptosis Detection Kit (Sangon Biotech).
Brie y, cells with different treatments were re-suspended with 100 μL binding buffer. Then cells were double stained by Annexin V for 15 min and propidium iodide (PI) for 10 min. Afterwards, the apoptotic rate was evaluated by using BD LSRII Flow Cytometer System (BD Biosciences) and the data was quantitated with the FACSDiva Software.
Brie y, cells with different treatments were re-suspended with 100 μL binding buffer. Then cells were double stained by Annexin V for 15 min and propidium iodide (PI) for 10 min. Afterwards, the apoptotic rate was evaluated by using BD LSRII Flow Cytometer System (BD Biosciences) and the data was quantitated with the FACSDiva Software.
9
Immunotherapy Conjugate Synthesis and Evaluation
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Fetal bovine serum was purchased from Wisent Biotechnology Co., Ltd. (Nanjing, China). Anti-PD-L1 (aPD-L1) was obtained from Invivogen (California, USA). The c(RGDyK) (cyclo(Arg-Gly-Asp-D-Tyr-Lys))
peptide and the Gly-PLGLAG-Cys polypeptide were synthesized by GL Biochem Co., Ltd. (Shanghai, China). 3-Maleimidobenzoic acid N-hydroxysuccinimide ester (MBS) and 2-Iminothiolane hydrochloride (Traut's Reagent) were purchased from Sigma-Aldrich (St. Louis, MO, USA). Annexin V-FITC Apoptosis Detection Kit was obtained from Sangon Biotech (Shanghai, China). Rabbit anti-caspase 9 antibody and anti-caspase 3 antibody were purchased from Boster Biological Engineering Co., Ltd. (Wuhan, China).
Rabbit anti-CD3 antibody was obtained from Proteintech (Wuhan, China), anti-CD4 antibody and anti-CD8 antibody were purchased from Abcam (Shanghai, China).
peptide and the Gly-PLGLAG-Cys polypeptide were synthesized by GL Biochem Co., Ltd. (Shanghai, China). 3-Maleimidobenzoic acid N-hydroxysuccinimide ester (MBS) and 2-Iminothiolane hydrochloride (Traut's Reagent) were purchased from Sigma-Aldrich (St. Louis, MO, USA). Annexin V-FITC Apoptosis Detection Kit was obtained from Sangon Biotech (Shanghai, China). Rabbit anti-caspase 9 antibody and anti-caspase 3 antibody were purchased from Boster Biological Engineering Co., Ltd. (Wuhan, China).
Rabbit anti-CD3 antibody was obtained from Proteintech (Wuhan, China), anti-CD4 antibody and anti-CD8 antibody were purchased from Abcam (Shanghai, China).
10
Apoptosis Detection via Flow Cytometry
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Cell-cycle distribution was detected by ow cytometry (FACScan, Becton 4 Dickinson). Apoptosis detection refers to the instruction manual of the Annexin V-FITC Apoptosis Detection Kit (Sangon, China). Treated cells were diluted into 1 × 10 6 cells/ml suspension and resuspended in 500 µl buffer solution. 5 µl PI and 5 µl Annexin V-FITC were added to incubate 30 min in the dark at room temperature. Analysis of apoptotic cells was performed by ow cytometry.
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