Cells were treated with indicated concentrations of PF-06651600 or DMSO for 48 h at 37°C. After incubation, cells were collected and washed twice with pre‐cold phosphate-buffered saline (PBS). About 6 × 105 cells were resuspended in 100 μl of 1× BD binding buffer solution (#556454, BD) and then stained with 7‐ADD (#559925, BD) and Annexin V‐PE (#556422, BD) in the dark for 15 min. Finally, 400 μl of 1× BD binding buffer solution was added to stop the dyeing. The cells were then measured on a Guava easyCyte flow cytometer (Merck, USA).
Binding buffer solution
Manufactured by BD
1X binding buffer solution is a laboratory reagent used to facilitate the binding of target molecules, such as proteins or nucleic acids, to a solid support or matrix during various experimental procedures. It provides the necessary chemical and ionic conditions for optimal binding interactions.
Automatically generated - may contain errors
Lab products found in correlation
Annexin 5 fluorescein isothiocyanate fitc apoptosis detection kit, by BD (2 mentions)
Accuri c6 software 1, by BD (2 mentions)
Guava easycyte flow cytometer, by Merck Group (1 mentions)
Annexin 5 pe, by BD (1 mentions)
7 add, by BD (1 mentions)
Facscalibur flow cytometer, by BD (1 mentions)
Cellquest pro software version 5, by BD (1 mentions)
Accutase cell detachment solution, by Thermo Fisher Scientific (1 mentions)
Accuri c6 flow cytometer, by BD (1 mentions)
Accuri c6, by BD (1 mentions)
Fitc annexin 5 apoptosis detection kit 1, by BD (1 mentions)
Annexin 5 fluorescein isothiocyanate, by BD (1 mentions)
Beckman fc500 flow cytometer, by Beckman Coulter (1 mentions)
5 protocols using binding buffer solution
1
Apoptosis Assay with PF-06651600
2
Propidium Iodide Viability Assay for BAE Cells
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Propidium iodide (PI) cell viability assay was used to assess the viability of BAE cells following treatment as indicated above. BAE cells were washed once with PBS and detached by incubation with accutase cell detachment solution (Thermo) at 37°C for 2 min. Both detached and floating cells were then collected and resuspended in 1X binding buffer solution (BD Biosciences) and stained with PI (BD Biosciences) for 15 min at room temperature in the dark. Samples were run on a FACSCalibur flow cytometer (BD Biosciences) and data were analyzed using CellQuest Pro software version 5.1 (BD Biosciences). BAE cells were identified by their forward-scatter (FSC) and side-scatter (SSC) properties. Viable and dead cell populations were identified as PI- and PI+ cells, respectively (18 (link)–20 (link)). A total of 10,000 single cell events were measured for each sample.
3
Annexin V-FITC Apoptosis Assay
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Cell apoptosis was examined using the Annexin V-fluorescein isothiocyanate (FITC) apoptosis detection kit (556547; BD Pharmingen, San Diego, CA, USA) according to the manufacturer’s instructions. Cells were re-suspended in 1X binding buffer solution (BD Pharmingen) with Annexin V-FITC and propidium iodide and incubated for 15 mins at room temperature in the dark. Apoptotic cells were analyzed using a BD Accuri C6 flow cytometer (BD Biosciences, Franklin Lakes, NJ, USA) with BD Accuri C6 software 1.0 (BD Biosciences).
4
Annexin V-FITC Apoptosis Assay for H460 Cells
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Cell apoptosis was examined using the Annexin V-fluorescein isothiocyanate (FITC) apoptosis detection kit (556547; BD Pharmingen, San Diego, CA, USA) according to the manufacturer's instructions. H460 cells were re-suspended in 1X binding buffer solution (BD Pharmingen) with Annexin V-FITC and propidium iodide (PI) and incubated for 15 min at room temperature in the dark. Apoptotic cells were analyzed using a BD Accuri C6 flow cytometer (BD Biosciences, Franklin Lakes, NJ, USA) with BD Accuri C6 software 1.0 (BD Biosciences).
5
Apoptosis Detection Using FITC Annexin V
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
FITC Annexin V Apoptosis Detection kit I (cat no. 556547; BD Biosciences) was used to assay cell apoptosis. Cultured cells, subsequent to treatment with pPeOp (30, 60 or 90 µg/ml), PVP (100 µg/ml) or 5-FU (100 µg/ml) under standard aforementioned conditions for 24 h, were collected with EDTA-free trypsin (0.25%) and washed twice with ice-cold PBS. Subsequently, cells (1×106/ml) were resuspended in 100 µl 1Xbinding buffer solution (BD Biosciences), to which 5 µl of fluorescein isothiocyanate/Annexin V (BD Biosciences) and 5 µl PI (BD Biosciences) were added. Cells were then gently vortexed and incubated for 15 min at room temperature (25°C) in the dark. Incubated cells were washed again with 1X binding buffer and resuspended in 200 µl of 1X binding buffer. PI (5 µl) and 400 µl of 1X binding buffer were added to each sample, and samples were analyzed using a Beckman FC500 flow cytometer (Beckman Coulter, Inc.). The software used to analyze apoptosis was CXP analysis (version 32; Beckman Coulter, Inc.).
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!