MSCs apoptosis was detected using the fluorescent dye PE Annexin V Apoptosis Detection Kit I (BD PharmingenTM, Franklin Lakes, NJ, USA, http://www.bd.com/ ) according to the manufacturer's instructions. Briefly, cells were rinsed with ice‐cold PBS and then resuspended in 200 μL of binding buffer. Five microlitres of Annexin V‐PE and 5 μL of 7‐AAD were added to the cells and incubated for 15 minutes at 25°C in the dark. Cells were analysed by FACS within 1‐hour. Ten thousands events were acquired on a FACSC‐LSR (Becton‐Dickinson, San Jose, CA, USA) and analysed with CellQuest (Becton‐Dickinson) software.
Facsc lsr
Manufactured by BD
Sourced in United States
The FACSC-LSR is a flow cytometry instrument designed for analyzing and sorting cells. It utilizes laser-based technology to detect and measure multiple characteristics of individual cells within a sample. The core function of the FACSC-LSR is to provide researchers and clinicians with data on cell size, granularity, and surface marker expression.
Automatically generated - may contain errors
Lab products found in correlation
Cellquest, by BD (3 mentions)
Annexin 5 fitc pi kit, by Merck Group (2 mentions)
Pe annexin 5 apoptosis detection kit 1, by BD (1 mentions)
Caspase 3 assay kit, by Takara Bio (1 mentions)
Fitc annexin 5 apoptosis detection kit 1, by BD (1 mentions)
Annexin 5 fitc pi apoptosis detection kit, by BD (1 mentions)
Trypsin, by Merck Group (1 mentions)
7 protocols using facsc lsr
1
Apoptosis Detection in MSCs
2
Apoptosis Determination in BM-MSCs
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
BM-MSC apoptosis was determined by flow cytometry with an Annexin V-FITC/PI Kit (Merck) according to the manufacturer’s instructions [21 (link)]. In brief, cells were re-suspended in 200 μL of binding buffer. Cells were incubated with 10 μL of Annexin V solution and 5 μL propidine iodide (PI) at room temperature for 30 min, respectively. The cells were immediately analyzed on a FACSC-LSR (Becton, Dickinson and Company, San Jose, CA). Meanwhile, the caspase-3 activity was measured using a Caspase-3 Assay kit (Clontech, MountainView, CA) according to the manufacturer’s instructions for 3 times. And these assays were performed in a blinded manner.
3
Quantifying Apoptosis by Flow Cytometry
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Apoptosis was assessed by flow cytometry using an FITC Annexin V Apoptosis Detection Kit I (556547, Becton Dickinson, Franklin Lakes, NJ, United States) according to the manufacturer’s instructions (Frey, 1997 (link)). Briefly, cells were rinsed with ice-cold PBS and then resuspended in 100 μL binding buffer (105 cells). A total of 5 μL Annexin V and 5 μL PI were added to each sample, and they were incubated for 15 min at 25°C in the dark. Then, 400 μL binding buffer was added to each tube and cells were immediately analyzed using a FACSC-LSR (Becton Dickinson) and evaluated with the Flow Jo 7.6 software.
4
Annexin V-FITC Apoptosis Assay for A549 Cells
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Fluorochrome-labeled Annexin V was used to specifically target and identify apoptotic cells. Fluorochrome-labeled Annexin V retains its high affinity for phosphatidylserine (PS) and thus serves as a sensitive probe for flow cytometric analysis of cells that are undergoing apoptosis. Since externalization of PS occurs in the earlier stages of apoptosis, FITC (Fluorescein isothiocyanate) Annexin V staining can identify apoptosis at an earlier stage than assays based on nuclear changes such as DNA fragmentation. PS exposure on the outer leaflet of the plasma membrane was detected using the Annexin V-FITC/PI Apoptosis Detection Kit (BD pharmingin TM, BD Biosciences Co., USA; Number 51–66121E) according to the manufacturer's instructions by flow cytometry. In brief, A549 cells were treated with either IC50 of the fungal crude extract, radiation, or IC50 of crude extract combined with radiation for 24 h. Untreated A549 cells were included as a control. Then, cells were rinsed with ice-cold PBS and then re-suspended in 500 μL of binding buffer. After full suspension, we added 5 μL of Annexin V-FITC and 10 μL of propidium iodide (PI). The cells were incubated for 5 min at 4 °C and then immediately analyzed on a FACSC-LSR (Becton, Dickinson and Company, San Jose, CA, USA) equipped with CellQuest (Becton, Dickinson and Company) software [36 (link)].
5
Measuring MSC Apoptosis by Flow Cytometry
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
MSC apoptosis induced by H/SD was determined by flow cytometry assay using an Annexin V-FITC/PI Kit (Merck, Whitehouse Station, New Jersey, USA) according to the manufacturer’s instructions [7 (link)]. Briefly, MSCs were harvested with 0.025% trypsin (Sigma Aldrich, Saint Louis, MO, USA) and incubated with 10 μl of Annexin V solution and 5 μl propidium iodide (PI) for 30 minutes at room temperature. The apoptosis of MSCs was analyzed on a FACSC-LSR (Becton, Dickinson and Company, San Jose, CA, USA).
6
Annexin V-FITC/PI Apoptosis Assay
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The Annexin V-FITC/PI Apoptosis Detection Kit was used to evaluate apoptosis of cells. After rinsed with cold PBS, cells were resuspended in 200 μl of binding buffer. Annexin V solution was added to the cells and incubated for 30 min at 4°C. The cells were then incubated with 5 ml propidium iodide (PI) and immediately analyzed with a FACScan. Ten thousand events were acquired on a FACSC-LSR (Becton-Dickinson, San Jose, CA) and analyzed with CellQuest (Becton-Dickinson) software.
7
Flow Cytometric Analysis of Apoptosis
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Apoptosis was detected by flow cytometric analysis. After transfection, the cells were washed twice with cold PBS and resuspended in 1X binding buffer at a concentration of 1 x 10 6 cells/mL. The cells were stained with annexin V and propidium iodide, using the annexin V apoptosis detection kit. After incubation at room temperature in the dark for 15 min, cell apoptosis was analyzed on an FACSC-LSR (Becton, Dickinson and Company, USA).
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!