MDA-MB-231 cells (1 × 106 cells/mL) were seeded in 6-well plates and treated with cinnamaldehyde (0, 10, 15, 20 μg/ml) for 24 h. A cell digestion solution (Beijing Solbio Technology Co., Ltd., article number: 20171024) was then used to prepare a cell suspension. Thereafter, 100 µl of the cell suspension was pipetted into a 1.5-ml Eppendorf tube. Subsequently, according to the instructions of the fluorescein thiocyanate (FITC)-conjugated Annexin-V apoptosis detection kit (Becton, Dickinson and Company, Franklin Lake, New Jersey), the cell suspension and 5 μl of Annexin-V-FITC were mixed with 5 μl of propidium iodide (PI, United States, batch number: 7040932) and incubated for 15 min. A 150-μl volume of the binding buffer was then added to each test tube and analyzed by flow cytometry.
Fitc conjugated annexin 5 apoptosis detection kit
Manufactured by BD
Sourced in United States
The FITC-conjugated Annexin V apoptosis detection kit is a laboratory product used to detect and quantify cells undergoing apoptosis or programmed cell death. The kit contains FITC-labeled Annexin V, a protein that binds to phosphatidylserine, which is exposed on the surface of apoptotic cells. This binding can be detected using flow cytometry or fluorescence microscopy.
Automatically generated - may contain errors
Lab products found in correlation
Facscalibur, by BD (4 mentions)
Lsrfortessa x 20 cell analyzer, by BD (3 mentions)
7 aminoactinomycin d 7 aad, by BD (2 mentions)
Propidium iodide, by BD (1 mentions)
Fitc conjugated anti cd24 and pe conjugated anti cd44 antibodies, by BD (1 mentions)
Fitc and pe conjugated anti mouse igg, by BD (1 mentions)
Thiazolyl blue tetrazolium bromide mtt powder, by Merck Group (1 mentions)
Stat3, by Cell Signaling Technology (1 mentions)
Gapdh, by Cell Signaling Technology (1 mentions)
12 well transwell plate, by Corning (1 mentions)
Edu medium, by RiboBio (1 mentions)
Dmi4000b, by Leica (1 mentions)
Apollo 567, by RiboBio (1 mentions)
Cellquest pro version 5, by BD (1 mentions)
Caspase glo assay kit, by Promega (1 mentions)
Varioskan lux multimode microplate reader, by Thermo Fisher Scientific (1 mentions)
Cck 8, by Dojindo Laboratories (1 mentions)
Lsrfortessa x 20 flow cytometer, by BD (1 mentions)
Facscalibur, by Beckman Coulter (1 mentions)
Lsrfortessa x 20, by BD (1 mentions)
21 protocols using fitc conjugated annexin 5 apoptosis detection kit
1
Cinnamaldehyde Induces Apoptosis in MDA-MB-231 Cells
2
Tetrandrine-Induced Apoptosis Assessment
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Cells were seeded in 6-well plates at a density of 1×106/well and treated with different concentrations of tetrandrine for 72h. Cells were harvested without EDTA-trypsin digestion solutions (Solarbio science & technology Co., Ltd., Beijing, lot. No: 20171024), washed twice with PBS, and resuspended in 200μl Binding Buffer, Pipette 100 μl of the cell suspension into Eppendorf tube 1.5ml). The cell suspension was mixed with 5μl of Annexin-V-fluorescein isothiocyanate (FITC) and 5μl Propidium Iodide (PI) according to the FITC conjugated Annexin-V apoptosis detection kit instructions (Becton, Dickinson and Company, Franklin Lake, New Jersey, USA, lot. No: 7040932), incubated for 15 min in the dark at room temperature. Then 150μl binding buffer was added to each tube and they were analyzed with a flow cytometer within 1 hour.
3
Apoptosis and Stem Cell Marker Analysis
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Cells were stained using a FITC-conjugated Annexin V apoptosis detection kit (BD Biosciences, Franklin Lakes, NJ, USA) according to the manufacturer’s protocol. For CD44+/CD24- staining analysis, cells were incubated for 30 min at 4°C with FITC- and PE-conjugated anti-mouse IgG or FITC-conjugated anti-CD24 and PE-conjugated anti-CD44 antibodies (BD Biosciences). Cells were analyzed by flow cytometry using a Beckman Coulter Expo.
4
Apoptosis Induction in Lung Cancer
5
Extraction and Analysis of Herbal Extract Compounds
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
SH003 was prepared as described in our previous study [26 (link)]. In brief, SH003 was provided from Hanpoong Pharm and Foods Company (Jeonju, Republic of Korea). Am, Ag, and TK were mixed at 1 : 1 : 1 ratio (w/w) and then extracted with 30% ethanol at 100°C for 3 h. Dried extracts were dissolved in 30% ethanol and stored at −80°C until use. DTX (Sigma-Aldrich, St. Louis, MO, USA) was dissolved in DMSO and stored at −20°C.
Thiazolyl blue tetrazolium bromide (MTT) powder was purchased from Sigma-Aldrich (St. Louis, MO, USA). FITC-conjugated Annexin V apoptosis Detection Kit and 7-aminoactinomycin D (7-AAD) were purchased from BD Pharmingen™ (BD Biosciences, San Jose, CA, USA) and Sigma-Aldrich (St. Louis, MO, USA), respectively. Anticleaved caspase-3, -PARP, -GAPDH, -p-EGFR (Tyr 1068), -p-EGFR (Tyr 1173), -EGFR, -p-AKT (Ser 473), -AKT, -p-C-Raf (Ser 338), -p-STAT3, and -STAT3 antibodies were purchased from Cell Signaling (Danvers, MA, USA). Anti-p-ERK and ERK antibodies were purchased from Santa Cruz Biotechnology (Santa Cruz, CA, USA). Anti-Ki67 and CD31 antibodies were purchased from Abcam (Cambridge, MA, USA). Horseradish peroxidase- (HRP-) conjugated secondary antibodies for mouse and rabbit were purchased from SeraCare Life Sciences (Milford, MA, USA).
Thiazolyl blue tetrazolium bromide (MTT) powder was purchased from Sigma-Aldrich (St. Louis, MO, USA). FITC-conjugated Annexin V apoptosis Detection Kit and 7-aminoactinomycin D (7-AAD) were purchased from BD Pharmingen™ (BD Biosciences, San Jose, CA, USA) and Sigma-Aldrich (St. Louis, MO, USA), respectively. Anticleaved caspase-3, -PARP, -GAPDH, -p-EGFR (Tyr 1068), -p-EGFR (Tyr 1173), -EGFR, -p-AKT (Ser 473), -AKT, -p-C-Raf (Ser 338), -p-STAT3, and -STAT3 antibodies were purchased from Cell Signaling (Danvers, MA, USA). Anti-p-ERK and ERK antibodies were purchased from Santa Cruz Biotechnology (Santa Cruz, CA, USA). Anti-Ki67 and CD31 antibodies were purchased from Abcam (Cambridge, MA, USA). Horseradish peroxidase- (HRP-) conjugated secondary antibodies for mouse and rabbit were purchased from SeraCare Life Sciences (Milford, MA, USA).
6
Microglia-Driven NSC Proliferation and Apoptosis
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
A total of 1 × 106 microglia were harvested and placed into the upper chamber of a 12-well trans-well plate (0.65 µm, Corning, New York, NY, USA), containing 1 µg/ml LPS. In addition, 1 × 106 NSCs were placed into the lower chamber, wells contained either culture medium; 40 nM leukosomes; or 40 nM lnc-EPS-leukosomes (n = 8). The microglia and medium in the control group were not untreated.
After culturing for 24 h at 37 °C, NSCs were cultured overnight using EdU medium (Ribobio, Guangzhou, China) and treated with 0.3% Triton X-100 in 1% fetal bovine serum (FBS) for 15 min at 37 °C. Then, cells were washed with PBS and to stain mitotic NSCs, 100 μL 1 × Apollo-567 (Ribobio, Guangzhou, China) was added to each well. DAPI was used to stain nuclei. Cells were evaluated and imaged using a fluorescence microscope (DMI4000B, Leica, Germany) at 565 nm and 461 nm.
The grouping and stimulation methods were identical as mentioned above. After 48 h, NSC were harvested, washed, and resuspended in PBS. A FITC-conjugated Annexin V Apoptosis Detection Kit (BD Biosciences, Franklin, NJ, USA) was used to identify apoptotic NSCs. Procedures were strictly performed by following the kit instructions, and cell apoptosis was evaluated by FACScan laser flow cytometer (CyFlow® Cube, Partec, Germany).
After culturing for 24 h at 37 °C, NSCs were cultured overnight using EdU medium (Ribobio, Guangzhou, China) and treated with 0.3% Triton X-100 in 1% fetal bovine serum (FBS) for 15 min at 37 °C. Then, cells were washed with PBS and to stain mitotic NSCs, 100 μL 1 × Apollo-567 (Ribobio, Guangzhou, China) was added to each well. DAPI was used to stain nuclei. Cells were evaluated and imaged using a fluorescence microscope (DMI4000B, Leica, Germany) at 565 nm and 461 nm.
The grouping and stimulation methods were identical as mentioned above. After 48 h, NSC were harvested, washed, and resuspended in PBS. A FITC-conjugated Annexin V Apoptosis Detection Kit (BD Biosciences, Franklin, NJ, USA) was used to identify apoptotic NSCs. Procedures were strictly performed by following the kit instructions, and cell apoptosis was evaluated by FACScan laser flow cytometer (CyFlow® Cube, Partec, Germany).
7
Apoptosis Analysis via Annexin V-FITC and 7-AAD
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Apoptosis was analyzed by the double-staining method using FITC-conjugated annexin V apoptosis detection kit and 7-amino-actinomycin D (7-AAD) purchased from BD PharmingenTM (BD Biosciences, San Jose, CA, USA) and Sigma-Aldrich, respectively. A total of 105 cells/well were cultured in a 6-well plate. Following a 24-h incubation at 37 °C, cells were treated with the considered concentration based on CI value and incubated for 48 h at 37 °C. Cells were collected and resuspended in 1× annexin binding buffer at a concentration of 1 × 105 cells/500 µL, followed by staining with annexin V-FITC or 7-AAD (BD 556570, BD Biosciences). Stained cells were analyzed using FACSCalibur (342973, BD FACSCalibur™, San Jose, CA, USA), and apoptotic cells were analyzed using Cell Quest Pro version 5.2 (BD Biosciences).
8
Apoptosis Analysis by Flow Cytometry
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The levels of cell apoptosis were analyzed by flow cytometry using the FITC conjugated Annexin V Apoptosis Detection Kit (BD Biosciences) following the manufacturer’s protocol. The cells in the 6-well plate were lysed with 0.25% trypsin, and then the cell pellets were collected by centrifugation (1500 RCF, 4 °C, 5 min). The cells were then washed twice by using cold PBS together with 1× binding buffer (1 × 105 cells/100 μL binding buffer) and were stained with annexin V-FITC and propidium iodide (PI) for 30 min at room temperature in the dark Each experiment was performed in triplicate.
9
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 with 95% ethanol containing 0.5% Tween-20 for 24 h, and incubated with propidium iodide (PI, 50 mg/mL) and RNase (50 mg/mL) for 30 min. For the Annexin V/PI assay, a FITC-conjugated Annexin V apoptosis detection kit (BD Biosciences, Franklin Lakes, NJ) was used according to the manufacturer’s protocol. Stained cells were analyzed by flow cytometry using BD LSRFortessaTM X-20 Cell Analyzer (BD Biosciences).
10
Caspase Activity and Apoptosis Assay
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
For measurement of caspase activity, HeLa-Parkin cells were transfected with EV or VZV gE and treated with 3 μM staurosporine for 2 h. A Caspase-Glo assay kit (Promega) was used according to the manufacturer’s instructions. Luminescence was measured using a Varioskan LUX Multimode Microplate Reader (Thermo Fisher Scientific). To measure apoptotic cell death, cells were harvested and stained using a fluorescein isothiocyanate (FITC)-conjugated Annexin V Apoptosis Detection Kit (BD Biosciences) according to the manufacturer’s instructions. The stained cells were analyzed using a BD LSR Fortessa X-20 flow cytometer (BD Biosciences). To examine cell cytotoxicity in CCCP-treated MRC5 and HaCaT cells, a CCK-8 (Dojindo, USA) assay was performed according to the manufacturer’s instructions.
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!