To detect cellular ROS, detached cells were loaded with CM-H2DCFDA (10 μM) for 10 min at 37 °C. Mitochondrial damage was measured in cells stained with either TMRE (50 nM) or NAO (50 nM) at 37 °C for 20 min. To analyze cell death, cells were resuspended in annexin binding buffer and labeled with annexin-V-FITC and propidium iodide (PI) at 25 °C for 15 min, according to the manufacturer’s instructions (Annexin-V-FITC and PI kits; BD Biosciences, San Jose, CA, USA). Cells were analyzed using a FACSCalibur flow cytometer (BD Biosciences, San Jose, CA, USA) with excitation wavelength 488 nm and observation wavelength 530 nm for green fluorescence and 585 nm for red fluorescence. Relative change in fluorescence was analyzed with FlowJo software 10.9 (FlowJo LLC, Ashland, OR, USA).
Annexin 5 fitc and pi kit
Manufactured by BD
Sourced in United States
The Annexin V-FITC and PI kit is a laboratory reagent used for the detection and quantification of apoptosis in cell samples. Annexin V is a protein that binds to phosphatidylserine, which is exposed on the surface of cells undergoing apoptosis. The kit includes Annexin V conjugated with the fluorescent dye FITC and the DNA-binding dye propidium iodide (PI). This allows for the identification of early apoptotic, late apoptotic, and necrotic cells through flow cytometry analysis.
Automatically generated - may contain errors
Lab products found in correlation
7 protocols using annexin 5 fitc and pi kit
1
Multiparametric Analysis of Cellular Stress
2
Mitochondrial Damage and Cell Death Analysis
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Mitochondrial damage was measured in cells stained with either TMRE (50 nM) or NAO (50 nM) at 37 °C for 20 min. To analyze cell death, cells were resuspended in annexin binding buffer and labeled with annexin-V-FITC and propidium iodide (PI) at 25 °C for 15 min, according to the manufacturer’s instructions (Annexin-V-FITC and PI kits; BD Biosciences, San Jose, CA, USA). Cells were analyzed using a FACSCalibur flow cytometer (BD Biosciences) with excitation wavelength 488 nm and observation wavelength 530 nm for green fluorescence and 585 nm for red fluorescence. Relative change in fluorescence was analyzed with FlowJo software 10.9.
3
Comprehensive Antibody and Reagent List
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The following primary antibodies were used: anti-LC3B (2775), anti-SQSTM1/p62 (5114), anti-β-Actin (4970), anti-COX2 (12282) and anti-LAMP1 (9091) were purchased from Cell Signaling Technology. Anti-DRP1 (sc271583), anti-NBR1 (sc-130380) and anti-MnSOD (sc-133254) were from Santa Cruz Biochemistry. Anti-COX1 (ab109025) was from Abcam. Goat secondary antibodies to rabbit IgG labelled with Alexa Fluor 546 (A11035), Alexa Fluor 488 (A11034), MitoSOX (M36008), dihydroethidium (DHE; D11347), MitoTracker Red (M7512), LysosensorGreen DND-189 (L7535), DQ-BSA Red (D12051), 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT, M6494), 5- (and-6)-chloromethyl-2′,7′-dichlorodihydrofluorescein diacetate (CM-H2DCFDA; C6827) were purchased from Thermo Fisher Scientific. Acetylsalicylic acid (Aspirin, A5376), allopurinol (A8003), diclofenac sodium (D6899) and Mito-Tempo (SML0737) were from Sigma Aldrich. ML171 (4653), and Mito-PY1(4428) were from Tocris Bioscience. Clioquinol (ClioQ; Santa Cruz Biotechnology, sc-201066), Magic Red Assay kit (Immuno Chemistry Technologies, 938), and Annexin-V-FITC and PI kits (BD Biosciences, 556547) were also used.
4
Apoptosis Analysis via Annexin V-FITC/PI
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Apoptosis analysis was performed using the Annexin V-FITC and PI kit (BD Biosciences) as per the manufacturer’s protocol. After staining, the cells were acquired in the MACSQuant Analyzer flow cytometer (Miltenyi Biotech).
5
Analyzing Apoptosis in KCL22 and K562 Cells
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The KCL22 and K562 cells were seeded into 6-well plates (5 × 105/well) and were both either transfected with LV-MEG3 or LV-control, or transfected with the miR-147 mimics or miR-147 control. Apoptosis was determined using an AnnexinV-FITC and PI kit (BD Biosciences, Franklin Lakes, NJ, USA) and was analyzed using a BD FACSCanto II system (BDBiosciences). The patient-derived cells were also assayed using the apoptosis kit after treatment with chidamide, as described above.
6
Apoptosis Analysis by Flow Cytometry
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Cells were harvested at 48 h after siRNA transfection or viral infection, washed with PBS, double stained with Annexin V-FITC and PI kit (BD Biosciences), and analyzed on a flow cytometer (BD Biosciences).
7
Annexin V/PI Apoptosis Assay
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The apoptosis assay of A549 and SPCA1 cells was conducted by Annexin V/FITC and PI kit (BD Biosciences, San José, CA). As mentioned previously50 (link), after washing with 0.01 M PBS twice, the cells were stained first with FITC-labeled Annexin-V for 30 min at room temperature and in the dark, and then with PI for 5 min. Finally, the stained cells were immediately tested by flow cytometer (BD Accuri™ C6 Plus). The flow cytometer collected 200,000 to 30,000 cells per test.
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!