Annexin V/propidium iodide (PI) assay was performed according to the manufacturer instructions (Annexin V-FITC kit; Roche Life Sciences, Basel, Switzerland). In brief, cells were washed with PBS, then incubated with the mixture of Annexin V and PI at room temperature for 10 min in the dark. The fluorescence signal was detected by using a fluorescence microscope (Olympus, Tokyo, Japan). Photomicrographs were obtained using identical exposure settings, and all images were quantified using Image-Pro Plus 6.0 software (Media Cybernetics, Rockville, MD, U.S.A.).
Annexin 5 fitc kit
Manufactured by Roche
Sourced in Switzerland
The Annexin V-FITC kit is a laboratory reagent used for the detection and quantification of apoptosis, or programmed cell death, in cell samples. The kit contains Annexin V conjugated to the fluorescent dye FITC, which binds to phosphatidylserine, a molecule that is exposed on the cell surface during the early stages of apoptosis. 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
Fluorescence microscope, by Olympus (1 mentions)
Image pro plus 6, by Media Cybernetics (1 mentions)
Facsaria, by BD (1 mentions)
Pb180325, by Procell (1 mentions)
Rnase a, by Merck Group (1 mentions)
Propidium iodide, by Merck Group (1 mentions)
Flow cytometry, by Beckman Coulter (1 mentions)
Expo32, by Beckman Coulter (1 mentions)
7 protocols using annexin 5 fitc kit
1
Annexin V and Propidium Iodide Staining
2
Apoptosis Quantification in HUVECs
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
After treated with 5 μg/mL ISMN for 24 h, adherent HUVECs were released by
trypsinization. The cell apoptosis was investigated by Annexin V: FITC kit
(Roche, Basel, Switzerland) based on the manufacturer’s specifications. The
samples were analyzed by a Becton Dickinson FACS Aria cell sorter.
trypsinization. The cell apoptosis was investigated by Annexin V: FITC kit
(Roche, Basel, Switzerland) based on the manufacturer’s specifications. The
samples were analyzed by a Becton Dickinson FACS Aria cell sorter.
3
Apoptosis Induction in HIF-Silenced Cells
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
SW480 control or HIF-1α or HIF-2α-silenced cells were seeded onto 24-well plates at a density of 1.5×105 cells per well. Twenty-four hours after seeding, the cells were incubated in the absence or presence of the apoptosis inducer H2O2 (1 mM) for 12 h. Apoptosis was measured by flow cytometry using the Annexin V FITC kit (Roche) as recommended by the manufacturer’s instructions. Necrosis was measured by propidium iodide (PI) permeability in the absence of detergent. The percentage of apoptotic cells was determined by flow cytometry.
4
Annexin V-FITC/PI Flow Cytometry Analysis
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The Annexin V-fluorescein isothiocyanate (FITC)/propidium iodide (PI) double staining method was used for cell apoptosis. Following 48 h of transfection, the cells were treated with 0.25% trypsin [without ethylenediaminetetraacetic acid (EDTA)], and the cells were collected in the flow tube, centrifuged at 4°C at 201 × g with the supernatant discarded. The cells were rinsed with cold PBS 3 times, and the supernatant was discarded. According to the instructions provided with the Annexin V-FITC kit (purchased from Roche, Basel, Switzerland), Annexin V-FITC/PI dying buffer was prepared by mixing Annexin V-FITC, PI, 4-(2-hydroxyethyl)-1-piper-azineethanesulfonic acid (HEPES) buffer solution at the proportion of 1:2:50. The cells were incubated at room temperature for 15 min, and 1 ml HEPES buffer solution (PB180325; Procell, Wuhan, China) was added, followed by shaking and evenly mixing the solution. The fluorescence of FITC and PI was detected by 525 and 620 nm bandpass filters at a wavelength of 488 nm by using a flow cytometer (LSR-II; BD Biosciences, Franklin Lakes, NJ, USA), and apoptosis was detected. The experiment was repeated 3 times.
5
Apoptosis Induction in ESCC Cells
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Apoptosis of ESCC cell spheres administrated 24 hours with SP (500 and 1000 nM)/aprepitant (30 μM) was assessed with the annexin V-FITC kit following manufacturer's directions (Roche Applied Science, Germany). Finally, flow cytometric analysis was performed on a flow cytometer (BD Bioscience, San Diego, CA, USA), and data analysis was assessed by FlowJo software (Treestar, OR, USA). All the treatments were conducted in triplicates.
6
Flow Cytometry Analysis of Apoptosis
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Cells were harvested and fixed in ethanol (70%) at the indicated time after incubation in the different conditions (control, TNF, PK 11195, and TNF + PK 11195). Cells were washed in PBS and incubated for 30 minutes at room temperature in the presence of RNAse A (100 μg/mL; Sigma-Aldrich Co.). Propidium iodide (PI; Sigma-Aldrich Co.) was added at a final concentration of 75 μg/mL and kept in the dark for 15 minutes at 37°C. PI fluorescence was measured by flow cytometry (Beckman Coulter Inc., Villepinte, France) with a laser excitation wavelength set at 620 nm and an emission pass band set at 488–540 nm.
Quantification of apoptosis was made using the AnnexinV–FITC kit (Roche Diagnostics, Meylan, France). Cells were grown in 12-well plates and harvested after the different treatments. Fluorescence was measured with the laser excitation wavelength set at 488 nm and an emission wavelength set at 518 nm using flow cytometry (Beckman Coulter Inc.). Quantitative analysis was performed using the EXPO 32 software (Beckman Coulter Inc.). PI was used as a control for the necrotic cells.
Quantification of apoptosis was made using the AnnexinV–FITC kit (Roche Diagnostics, Meylan, France). Cells were grown in 12-well plates and harvested after the different treatments. Fluorescence was measured with the laser excitation wavelength set at 488 nm and an emission wavelength set at 518 nm using flow cytometry (Beckman Coulter Inc.). Quantitative analysis was performed using the EXPO 32 software (Beckman Coulter Inc.). PI was used as a control for the necrotic cells.
7
Apoptosis Induction by 5'-FU and Adenovirus
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Apoptosis induced by 5'-FU treatment and adenovirus infection for 24 or 48 h was determined by Annexin V-FITC kit (Roche Diagnostics, Indianapolis, IN, USA). Cells were incubated in the dark with Annexin V-FITC and PI for 20 min. The apoptotic cells were quantified using flow cytometry, and the percentage of Annexin V-positive cells (early apoptosis) or Annexin V-plus-PI positive cells (late apoptosis) were calculated and were presented as total apoptotic cells. Caspase-3 activity was measured by Apo-ONE homogeneous caspase-3 assay (Promega, Madison, wI, USA) and was calculated and compared with control cells.
About PubCompare
Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.
We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.
However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.
Ready to get started?
Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required
Revolutionizing how scientists
search and build protocols!