K562 cells were treated with various concentrations of molecule L20 and SAHA for 24 h, cells were harvested and PBS washed twice, then resuspended with binding buffer (Becton Dickinson, USA). Cells were incubated with Annexin V-BV421 (Becton Dickinson, USA) and 7-AAD (Becton Dickinson, USA) double labelling for 30 min in the dark at room temperature and measured by flow cytometry (FACSAriaIII, Becton Dickinson, USA). The data was analysed using Flowjo-V10 software.
Annexin 5 bv421
Manufactured by BD
Sourced in United States
Annexin V BV421 is a fluorescently labeled protein that binds to phosphatidylserine, a component of the cell membrane. This binding can be used to detect and quantify apoptotic cells.
Automatically generated - may contain errors
Lab products found in correlation
Binding buffer, by BD (4 mentions)
Facsaria 3, by BD (3 mentions)
7 aad, by BD (3 mentions)
Fortessa flow cytometer, by BD (3 mentions)
Canto 2 special order research product, by BD (2 mentions)
Pi rnase a, by Abcam (2 mentions)
Lsrfortessa x 20 system, by BD (2 mentions)
Annexin binding buffer, by Thermo Fisher Scientific (2 mentions)
Celltracker deep red, by Thermo Fisher Scientific (2 mentions)
Sytox blue, by Thermo Fisher Scientific (2 mentions)
Fitc conjugated cd71 okt9, by Thermo Fisher Scientific (2 mentions)
Apc conjugated cd49d 9f10, by BioLegend (2 mentions)
Sytox orange, by Thermo Fisher Scientific (2 mentions)
Facscanto 2, by BD (2 mentions)
Arachidonic acid, by Helena Laboratories (1 mentions)
Zombie green fixable viability dye, by BioLegend (1 mentions)
Mitospy green fm, by BioLegend (1 mentions)
Mitosox red, by Thermo Fisher Scientific (1 mentions)
Lsrfortessa, by BD (1 mentions)
Annexin buffer, by BD (1 mentions)
20 protocols using annexin 5 bv421
1
Annexin V-BV421 and 7-AAD Assay for Apoptosis
2
Multicolor Flow Cytometry of Platelets
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
BRT was purchased from Frontier Scientific Inc. (Logan, UT, USA). All phlebotomy consumables, Annexin V Binding Buffer, Stain Buffer (BSA), Compensation Beads (anti-Mouse Ig, κ/Negative Control) and anti-CD42b-APC (HIP1, 551061), anti-CD42b-PE-Cy5 (HIP1, 551141), anti-CD62P-PE (AK4 555524), anti-PAC-1-FITC (PAC1, 340507) and Annexin V-BV421 (563973) were purchased from Becton Dickinson (Brisbane, Australia). Platelet agonists adenosine diphosphate (ADP), collagen and arachidonic acid (AA) were purchased from Helena Laboratories (Melbourne, Australia) with thrombin receptor activating peptide SFLLRN (TRAP-6) purchased from Haemoview Diagnostics (Brisbane Australia). CHRONO-LUME® and all aggregation consumables were purchased from DKSH Australia (Brisbane, Australia) with MitoSOX™ Red from ThermoFisher Scientific (Brisbane, Australia). Both MitoSPY™ Green FM and Zombie Green™ Fixable Viability Dye were purchased from BioLegend (San Diego, USA). All other reagents were purchased from Sigma Aldrich (Castle Hill, Australia) unless otherwise stated.
3
Apoptosis Induction in K562 Cells
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
K562 cells were treated with various concentrations of molecule D28 and SAHA for 24 h, cells were harvested and PBS washed twice, then resuspended with binding buffer (Becton Dickinson, United States). Cells were incubated with Annexin V-BV421(Becton Dickinson, United States) and 7-AAD (Becton Dickinson, United States) double labeling for 30 min in the dark at room temperature and measured by flow cytometry (FACSAriaIII, Becton Dickinson, United States). The data was analyzed by using Flowjo-V10 software.
4
Apoptosis Induction in K562 Cells
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
K562 cells were treated with various concentrations of molecule C2 , olaparib, or chlorambucil for 24 h. Cells were then harvested and washed twice with PBS and then resuspended with binding buffer (Becton Dickinson, United States). Cells were incubated with Annexin V-BV421 (Becton Dickinson, United States) and 7-AAD (Becton Dickinson, United States) for double labeling for 30 min, in the dark, at room temperature, followed by analysis using flow cytometry (FACSAria III, Becton Dickinson, United States). The data were analyzed using FlowJo v10 software.
5
Annexin V Staining for Cell Viability
6
Annexin V Apoptosis Assay by Flow Cytometry
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
For Annexin V labeling, cells were washed once with PBS and resuspended in 50 µL Binding Buffer (BD Biosciences, France) in which 2.5 µL of AnnexinV BV421 (BD Biosciences, France) and 2.5 µL of PI (Sigma-Aldrich, Saint-Louis, MO, USA) at 0.5 mg/mL were added. Cells were incubated 15 min at room temperature in the dark before adding 200 µL of Binding Buffer. The cells were analyzed by flow cytometry using 488 nm excitation and measuring violet fluorescence emission for Annexin V (450/50 bandpass) and red fluorescence emission for PI (610/20 bandpass) on the BD LSRFortessa™ cell analyzer (BD Biosciences, France). Data were exported and analyzed with Flowjo software for evaluation of the percentage of Annexin V-positive cells and of the FSC median.
7
High-Sensitivity Flow Cytometry of Apoptotic Cells
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
All the analyses were performed on a BD Canto II Special Order Research Product (BD Biosciences) equipped with a small particle option, as described previously.22 (link),23 (link) Perfusates (10 µL) were labeled in a total reaction volume of 50 μL at 32°C for 60 min with LWA 300 probe (300 nM). Cell tracker deep red (1 µM, ThermoFicher Scientific) was added for 30 min at 32°C and then 1 µL of Annexin V-BV421 (BD Biosciences) was added for 20 min at room temperature. Then, the sample was diluted by adding 250 µL of labeling buffer prior analysis by high-sensitivity flow cytometry.
8
Apoptosis and Necrosis Quantification in U-87 Cells
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The induction of apoptosis and necrosis
of U-87 was measured 4 h after X-ray exposure by flow cytometry. X-ray-exposed
unstained and NT-PEO-Por*-stained U-87 were detached from dishes by
0.25% trypsin–EDTA, centrifuged at 1200 rpm for 5 min, and
resuspended in 100 μL of the annexin buffer (BD) and 5 μL
of Annexin V BV421 (BD), which binds to phosphatidylserin (PS) residues
and allows the identification of apoptotic cells. Cells were incubated
a 4 °C for 20 min, washed in the annexin buffer, and centrifuged.
The viability dye 7-aminoactinomycin D (7-AAD, BD) was added to cell
pellets to stain non-viable cells and recognize late-apoptotic fractions.
For fluorescence-activated cell sorting (FACS) characterization, data
were obtained using a FACSAria Fusion cell sorter, equipped with five
lasers, and analyzed with FACSDiva software (ver. 8.0, BD). At least
10 × 103 events were recorded for each condition.
Debris events were excluded from the analysis by morphological gating
(side scatter vs forward scatter dot plot).
of U-87 was measured 4 h after X-ray exposure by flow cytometry. X-ray-exposed
unstained and NT-PEO-Por*-stained U-87 were detached from dishes by
0.25% trypsin–EDTA, centrifuged at 1200 rpm for 5 min, and
resuspended in 100 μL of the annexin buffer (BD) and 5 μL
of Annexin V BV421 (BD), which binds to phosphatidylserin (PS) residues
and allows the identification of apoptotic cells. Cells were incubated
a 4 °C for 20 min, washed in the annexin buffer, and centrifuged.
The viability dye 7-aminoactinomycin D (7-AAD, BD) was added to cell
pellets to stain non-viable cells and recognize late-apoptotic fractions.
For fluorescence-activated cell sorting (FACS) characterization, data
were obtained using a FACSAria Fusion cell sorter, equipped with five
lasers, and analyzed with FACSDiva software (ver. 8.0, BD). At least
10 × 103 events were recorded for each condition.
Debris events were excluded from the analysis by morphological gating
(side scatter vs forward scatter dot plot).
9
Cell Cycle and Apoptosis Analysis
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Cells were harvested by centrifugation and washed in cold phosphate‐buffered saline (PBS). For cell cycle analysis and enumeration of polyploid cells, cells were fixed with ice‐cold 66% ethanol overnight at 4 °C and stained with propidium iodide (PI)/RNase A (Abcam) for 30 min at 37 °C in the dark. For Annexin V/PI quantification, cell were resuspended in 1× Annexin binding buffer (Thermo Fisher Scientific) and stained with Annexin V BV421 (BD Biosciences, Franklin Lakes, NJ, USA) and PI at room temperature for 15 min. Flow cytometry analyses were performed using the LSRFortessa X‐20 system and flowjo software (BD Biosciences).
10
Multicolor Flow Cytometry Analysis
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
We performed FACS analysis of CD11b, CD14 and CD15 expression using the following anti-human monoclonal antibodies: CD11b-PE (Beckman Coulter, IM2581U), CD14-AF700 (BD, 557923), CD15-APC (BD, 551376), CD45-BV510 (BD, 563204) or CD45-APCH7 (BD, 641417). Apoptotic cells were stained with AnnexinV-BV421 (BD, 563973) or Annexin-FITC (BD, 556420) according to the manufacturer’s protocol (BD).
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!