Log In Sign up
The largest database of trusted experimental protocols

Annexin 5 pe

Manufactured by BioLegend
Visit Supplier
Sourced in United States

Annexin V-PE is a fluorescently labeled protein that binds to phosphatidylserine, a cell surface marker for apoptosis. It is used in flow cytometry applications to detect and quantify apoptotic cells.

Automatically generated - may contain errors

Lab products found in correlation

Annexin 5 binding buffer, by BioLegend (8 mentions) 7 aad, by BioLegend (8 mentions) Facscanto 2, by BD (3 mentions) 7 aad, by Thermo Fisher Scientific (3 mentions) 7 aad, by BD (3 mentions) Lsrfortessa, by BD (3 mentions) 4 6 diamidino 2 phenylindole (dapi), by Thermo Fisher Scientific (2 mentions) 7 aminoactinomycin d 7 aad, by BioLegend (2 mentions) 7 aad viability staining solution, by BioLegend (2 mentions) Attune flow cytometer, by Thermo Fisher Scientific (2 mentions) Annexin 5 binding buffer, by Thermo Fisher Scientific (2 mentions) Apc brdu flow kit, by BD (2 mentions) 9eg7 antibody, by BD (2 mentions) Facscanto 2 analyzer, by BD (2 mentions) Lsrfortessa cell analyzer, by BD (2 mentions) Facscalibur flow cytometer, by BD (2 mentions) Facsaria 2, by BD (2 mentions) Annexin 5 binding buffer, by BD (2 mentions) Alexa fluor 488, by Thermo Fisher Scientific (2 mentions) Lsrii flow cytometer, by BD (2 mentions)

Spelling variants of this product

Annexin V (328 citations) PE Annexin V Apoptosis Detection Kit with 7-AAD (22 citations) PE Annexin V Apoptosis Detection Kit (18 citations) PE conjugated Annexin-V (4 citations) Annexin V-PE/7-AAD (3 citations) PE-conjugated anti-Annexin V (1 citations)

55 protocols using annexin 5 pe

1

Apoptosis Profiling of B-ALL Cells

Check if the same lab product or an alternative is used in the 5 most similar protocols
Following 24 hours or 72 hours treatment of DMSO or SF2535, B-ALL cells were resuspended in 1X Annexin V binding buffer (Becton Dickinson) at a concentration of 1×106 cells per mL. 2.5μl Annexin V PE (BioLegend) and 2.5μl DAPI (50μg/mL, Invitrogen) were added to 100μl of the cell suspension. After 15 min incubation at room temperature in the dark, B-ALL cells were analyzed by flow cytometry using BD FACS Canto II.
Ruan Y., Kim H.N., Ogana H.A., Wan Z., Hurwitz S., Nichols C., Abdel-Azim N., Coba A., Seo S., Loh Y.H., Gang E.J., Abdel-Azim H., Hsieh C.L., Lieber M.R., Parekh C., Pal D., Bhojwani D., Durden D.L, & Kim Y.M. (2021). Preclinical Evaluation of a Novel Dual Targeting PI3Kδ/BRD4 Inhibitor, SF2535, in B-Cell Acute Lymphoblastic Leukemia. Frontiers in Oncology, 11, 766888.
+ Open protocol
+ Expand
2

NK Cell-Mediated Cytotoxicity Assay

Check if the same lab product or an alternative is used in the 5 most similar protocols
Target YAC‐1 cells were labelled with carboxyfluorescein succinimidyl ester (CFSE; C1031, Beyotime) at a final concentration of 5 μM. They were then co‐cultured with effector NK cells at an effector‐to‐target cell (E:T) ratio of 2:1 or 0:1 for 4 h at 37°C under 5% CO2. After 4 h, the samples were stained with Helix NP NIR (425301, BioLegend) and Annexin V‐PE (640908, BioLegend) for 10 min. Fluorescence‐activated cell sorting (FACS) analysis was performed immediately with a BD LSR Fortessa Cell Analyzer (BD Biosciences). The Incucyte S3 Live‐Cell Analysis Instrument (Sartorius AG) was used to monitor the killing activity of NK cells. Propidium iodide (C1052, Beyotime) was used to mark dead cells.
Li D., Ji W., Wei D., Gu A., Song Z., Fang W., Meng C., Yang Y, & Peng J. (2022). Cytochrome P450 26A1 regulates the clusters and killing activity of NK cells during the peri‐implantation period. Journal of Cellular and Molecular Medicine, 26(8), 2438-2450.
+ Open protocol
+ Expand
3

Assessing Treg Cell Apoptosis

Check if the same lab product or an alternative is used in the 5 most similar protocols
CD4+ T cells isolated from Foxp3Cre and Foxp3CremiR-142fl/fl spleens by EasySep Mouse CD4+ T Cell Isolation Kit (STEMCELL Technologies, Vancouver, British Columbia, Canada) were sorted for YFP expressing cells using BD FACSAria II instrument. CD4+YFP+ Treg cells were stimulated with anti-CD3 (1 μg/mL) specific antibodies for 48 hours, stained with Annexin V-PE (BioLegend) and analyzed by flow cytometry. For the IFNγ-induced apoptosis of Treg cells assay, splenocytes were cultured for 24 hours in the presence or absence of IFNγ (10 ng/mL) and then harvested and stained with Annexin V-PE for fluorescence activated cell sorting (FACS) analysis.
Wang W.L., Ouyang C., Graham N.M., Zhang Y., Cassady K., Reyes E.Y., Xiong M., Davis A.M., Tang K., Zeng D, & Boldin M.P. (2022). microRNA-142 guards against autoimmunity by controlling Treg cell homeostasis and function. PLoS Biology, 20(2), e3001552.
+ Open protocol
+ Expand
4

Annexin V and 7-AAD Staining

Check if the same lab product or an alternative is used in the 5 most similar protocols
Single cell suspensions were washed once with Hank’s Salt solution, containing Calcium and Magnesium (HSS) (Millipore, L 2035) and supplemented with 2% FCS. Cells were resuspended in 100μl of wash buffer containing the manufacturers recommended concentration of AnnexinV-PE (Biolegend, 640908). Samples were incubated for 20 minutes at 4°C along with surface marker antibody stains. Cells were then washed with 800μl of HSS plus 2% FCS and resuspended in 200μl. 7-AAD (Biolegend, 420404) was added to the 200μl cell suspension shortly before analysis on the flow cytometer at the manufacturers recommended concentration. Live cells were considered 7-AAD- and AnnexinV-.
Olson W.J., Jakic B., Labi V., Schoeler K., Kind M., Klepsch V., Baier G, & Hermann-Kleiter N. (2019). Orphan Nuclear Receptor NR2F6 Suppresses T Follicular Helper Cell Accumulation through Regulation of IL-21. Cell Reports, 28(11), 2878-2891.e5.
+ Open protocol
+ Expand
5

Flow Cytometric Analysis of Plasma Microvesicles

Check if the same lab product or an alternative is used in the 5 most similar protocols
Plasma samples were stained with the following antibodies: CD41a-FITC (BD Pharmingen), annexinV-PE (BioLegend, San Diego, CA, USA), CD45-BrilliantViolet421 (BioLegend), and CD144-PE/Cy7 (BioLegend) (1 µL of each per 50 µL of plasma) for 15 min at 37 °C, then fixed with PFA and diluted with PBS directly before the measurement. PFA and PBS were filtered through a 0.1 µm filter to reduce the background. Samples were measured on a NovoCyte Quanteon Flow Cytometer (Agilent, Santa Clara, CA, USA), flow rate slow, with no gating, for a fixed amount of time. Obtained data are presented as either densities (events/µL) or median florescence intensity (MFI). A flow cytometry schema of gating platelet-derived and procoagulant microvesicles is shown in Figure 1A.
Marcinkowska A., Wolska N., Luzak B., Cisiecki S., Marcinkowski K, & Rozalski M. (2022). Platelet-Derived Procoagulant Microvesicles Are Elevated in Patients with Retinal Vein Occlusion (RVO). Journal of Clinical Medicine, 11(17), 5099.
+ Open protocol
+ Expand
6

Evaluating T Cell Activation and Death

Check if the same lab product or an alternative is used in the 5 most similar protocols
To examine RICD, activated T cells (105 cells/well) were plated in triplicate in 96-well round-bottom plates and treated with anti-CD3ε mAb OKT3 (1–100 ng/ml) in cRPMI + 100 IU/ml rhIL-2 for 24 hours. R59949 (5–10 μM), R59022 (5–10 μM), DAG (50 μM), U0126 (5 μM), FR180204 (10 μM) or Rottlerin (6 μM) inhibitors were added 30 minutes before restimulation. At 24 hours post-restimulation, cells were stained with 1 μg/ml propidium iodide and collected for 30 seconds per sample on FACScan or Accuri C6 flow cytometers (BD). Cell death was analyzed with CellQuest/CFlow software (BD) or Flowing software (Turku Bioimaging) as percentage of cell loss = (1 − [number of viable cells (treated) / number of viable cells (untreated)]) × 100 (5 (link)).
For AnnexinV assays, ~1×106 cells were treated with OKT3 (10 ng/ml) as above. Cells were stained 6–12 hours later with AnnexinV-PE (Biolegend) and analyzed on an Accuri C6.
To evaluate CD25 expression, 1×106 cells were stimulated with OKT3 (100 ng/ml) for 24 hours, fixed and stained with anti-CD25 plus anti-mouse AlexaFlour488. Stained cells were collected on a FACSCalibur.
Ruffo E., Malacarne V., Larsen S.E., Das R., Patrussi L., Wülfing C., Biskup C., Kapnick S.M., Verbist K., Tedrick P., Schwartzberg P.L., Baldari C.T., Rubio I., Nichols K.E., Snow A.L., Baldanzi G, & Graziani A. (2016). Inhibition of diacylglycerol kinase alpha restores restimulation-induced cell death and reduces immunopathology in XLP-1. Science translational medicine, 8(321), 321ra7.
+ Open protocol
+ Expand
7

Apoptosis Assessment of ALL Cells

Check if the same lab product or an alternative is used in the 5 most similar protocols
The viability of the ALL cells after VDL, Tysabri, or P5G10 treatment was determined by Annexin V-PE (BioLegend, San Diego, CA, USA) and 7-aminoactinomycin D (7-AAD) (BioLegend) staining and flow cytometry (FACScalibur, BD Biosciences, San Diego, CA, USA). The FlowJo 7.6.5 software (FlowJo, LLC, Franklin Lakes, NJ, USA was used to analyze the acquired data.
Liu H.C., Gang E.J., Kim H.N., Ruan Y., Ogana H., Wan Z., Bönig H., Shung K.K, & Kim Y.M. (2020). Characterizing the Motility of Chemotherapeutics-Treated Acute Lymphoblastic Leukemia Cells by Time-Lapse Imaging. Cells, 9(6), 1470.
+ Open protocol
+ Expand
8

BCG-Induced RAW264.7-MptpB Cell Apoptosis

Check if the same lab product or an alternative is used in the 5 most similar protocols
RAW264.7-MptpB cells stimulated by BCG (MOI = 10) for 24 h were stained with 7AAD-PerCP-Cy5.5 and Annexin V-PE (Biolegend). The FACS Canto II flow cytometer of FACS Diva software was used to analyze the stained cells.
Fan L., Wu X., Jin C., Li F., Xiong S, & Dong Y. (2018). MptpB Promotes Mycobacteria Survival by Inhibiting the Expression of Inflammatory Mediators and Cell Apoptosis in Macrophages. Frontiers in Cellular and Infection Microbiology, 8, 171.
+ Open protocol
+ Expand
9

Apoptosis and Cell Cycle Analysis

Check if the same lab product or an alternative is used in the 5 most similar protocols
For apoptosis analysis, 1 × 105 cells were co-stained with annexin V-PE and 7-amino-actinomycin D (7-AAD, both from Biolegend, San Diego, CA, USA). Cell death was measured by a Flow Cytometer FACSCalibur (BD Biosciences, San Jose, CA, USA) as described before.46 (link) To determine cell-cycle distribution, 1 × 106 cells were fixed and stained with 70% cold ethanol and propidium iodide (PI). DNA contents were measured by flow cytometry. Data were analyzed using ModFit cell-cycle analysis software (Verity Software House).
Guan H., Tan P., Xie L., Mi B., Fang Z., Li J., Yue J., Liao H, & Li F. (2015). FOXO1 inhibits osteosarcoma oncogenesis via Wnt/β-catenin pathway suppression. Oncogenesis, 4(9), e166-.
+ Open protocol
+ Expand
10

T Cell Apoptosis Induction Assay

Check if the same lab product or an alternative is used in the 5 most similar protocols
Jurkat T cells or primary CD4+ T cells were cultured at 1 × 106 cells/ml for 16 h in RPMI 10% FCS, 20 μg/ml DNase. Anti-Apo-1 (anti-Fas) and Protein A (0.1-1 μg/ml, gift from Min-Li Weber) were added and the culture was continued for a further 5 h. Samples were washed in PBS and stained with live/dead blue stain. Cells were washed once in AnnexinV binding buffer (Biolegend) then stained with anti-CD4-PeCy5 (Beckman Coulter) and AnnexinV-PE (Biolegend) for 20 min at RT. The cells were washed and fixed for 20 minutes with 2% paraformaldehyde in AnnexinV binding buffer. Tax staining was performed as described above. Gating strategy is outlines in Additional file 8.
Rowan A.G., Suemori K., Fujiwara H., Yasukawa M., Tanaka Y., Taylor G.P, & Bangham C.R. (2014). Cytotoxic T lymphocyte lysis of HTLV-1 infected cells is limited by weak HBZ protein expression, but non-specifically enhanced on induction of Tax expression. Retrovirology, 11, 116.
+ Open protocol
+ Expand

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

Sign up now

Revolutionizing how scientists
search and build protocols!