MSCs were harvested and resuspended at the density of 2 × 105 cells/mL with the annexin-binding buffer. Then, 5 µL Annexin V Conjugate (Invitrogen; cat. no. A35110) was added to 100 μL of each cell suspension. Then, 400 µL annexin-binding buffer was added and the stained cells were analyzed using an ARIAII flow cytometer (BD) followed by analysis with FlowJo 10.8.1 (https://www.flowjo.com/ ).
Annexin 5 conjugate
Manufactured by Thermo Fisher Scientific
Annexin V Conjugate is a laboratory reagent used for the detection and quantification of apoptotic cells. It binds to phosphatidylserine, a phospholipid that is exposed on the surface of cells undergoing programmed cell death. The conjugate is typically labeled with a fluorescent dye, allowing for the identification and analysis of apoptotic cells using flow cytometry or other imaging techniques.
Automatically generated - may contain errors
Lab products found in correlation
Propidium iodide, by Thermo Fisher Scientific (1 mentions)
Alexa fluor 350 conjugate, by Thermo Fisher Scientific (1 mentions)
Lsr 2, by BD (1 mentions)
Ionomycin from streptomyces conglobatus, by Merck Group (1 mentions)
Facscalibur system, by BD (1 mentions)
Bz 9000, by Keyence (1 mentions)
Facs scanner, by BD (1 mentions)
Thiazolyl blue, by Merck Group (1 mentions)
6 protocols using annexin 5 conjugate
1
Annexin V-Based Apoptosis Assay
2
Annexin V Apoptosis Assay for Vemurafenib
3
Annexin V Staining for Erythrocyte Apoptosis
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Detection of erythrocyte apoptosis with Annexin V conjugates by flow cytometry (Annexin V conjugate, ThermoFisher scientific) was performed in a BD FACSCalibur™ system (BD Biosciences, San Jose, CA). Erythrocytes previously treated with ionomycin from Streptomyces conglobatus (Sigma-Aldrich) were used as a positive control for erythrocyte apoptosis. Erythrocytes derived from healthy animal blood with and without annexin were used as negative controls.
4
Cell Proliferation and Apoptosis Assays
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Cell proliferation was evaluated by measuring sequential cell viability using the Premix WST-8 Cell Proliferation assay kit (Dojindo Inc., Kumamoto, Japan) as described previously. 16 Cancer cell lines (5×10 3 cells/well each) were seeded into 96-well plates. The OD of the solution at 450 nm in each well was measured 2 hours after the addition of 10 μL of WST-8, and the fold-change in OD from the day of seeding (day 0) was evaluated. The assay was done in a technical quintuplicate.
The detection of apoptotic status was evaluated with annexin-V staining as described previously. 17 To be brief, cells are diluted in annexin-binding buffer and 10 µL of annexin-V conjugate (Thermo Fisher Scientific). The cells were mounted on slides, and the apoptotic cells labeled fluorescently were counted under ×200 magnification on BZ9000 (Keyence, Osaka, Japan). UVirradiated cells for 8h followed by 16h incubation in the normal condition was used as positive control.
The detection of apoptotic status was evaluated with annexin-V staining as described previously. 17 To be brief, cells are diluted in annexin-binding buffer and 10 µL of annexin-V conjugate (Thermo Fisher Scientific). The cells were mounted on slides, and the apoptotic cells labeled fluorescently were counted under ×200 magnification on BZ9000 (Keyence, Osaka, Japan). UVirradiated cells for 8h followed by 16h incubation in the normal condition was used as positive control.
5
Evaluating Cell Viability via Flow Cytometry
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The effect of NAP9 and NAPSC on cell viability was determined by flow cytometry (FACS scanner, Becton Dickinson) using propidium iodide (RD systems) and annexin V conjugates (Life Technologies), as provided by the manufacturers.
6
Cell Viability and Apoptosis Assays
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Cells were seeded at a density of 2–3 × 104 cells per well in 24-well plates and incubated at 37 °C in humidified 5% CO2 for 24 hours. Then the indicated drugs were added to the cells for the designated time and concentration. Cell viability was assessed by methylthiazolydiphenyl-tetrazolium bromide (MTT) assay. A 0.5-mg/mL solution of Thiazolyl Blue (Sigma) in phenol-free DMEM was added to cells at 37 °C for 1 hour. The substrate was then dissolved in isopropanol and absorbance was measured with a spectrophotometer at 570 nm. All MTT assays were performed 3 times in triplicate.
Cell apoptosis after indicated drug treatment was detected by flow cytometry with Annexin V conjugates following manufacturer’s instructions (Life Technologies).
Cell apoptosis after indicated drug treatment was detected by flow cytometry with Annexin V conjugates following manufacturer’s instructions (Life Technologies).
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!