Annexin 5 af568

Manufactured by Thermo Fisher Scientific

Sourced in United States

Annexin V-AF568 is a fluorescently labeled protein used to detect and quantify apoptotic cells. It binds to phosphatidylserine, a marker of apoptosis, and can be detected using flow cytometry or fluorescence microscopy.

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Lab products found in correlation

Hoechst 33342, by Thermo Fisher Scientific (1 mentions) Phorbol 12 myristate 13 acetate, by Merck Group (1 mentions) Gapdh, by Novus Biologicals (1 mentions) O phospho l serine, by Merck Group (1 mentions) Marimastat, by Bio-Techne (1 mentions) Gi254023x, by Bio-Techne (1 mentions) Ultraview vox, by PerkinElmer (1 mentions) Calcien am, by Thermo Fisher Scientific (1 mentions)

Close spelling variants of this product

Annexin V (493 citations) AF568 (12 citations)

2 protocols using annexin 5 af568

Annexin V-AF568 and Hoechst 33342 Staining

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Annexin V-AF568 and Hoechst 33342 were purchased from Thermo Fisher Scientific (Waltham, MA, USA). O-phospho-L-serine (OPS) and Phorbol 12-myristate-13-acetate (PMA) were obtained from Sigma Aldrich (St. Louis, MO, USA). Ionomycin (IO) was purchased from Merck Millipore (Darmstadt, Germany). Hydroxamate-based ADAM17/ADAM10 inhibitor GW280264X [30 (link)] was purchased from Aeobious (Gloucester, MA, USA). Marimastat and ADAM10 inhibitor GI254023X [31 (link)] were purchased from Tocris Bioscience (Bristol, UK). Additional antibodies used: ANO1 (Bio-Techne, Abingdon, UK), ANO4 (Aviva Systems Biology, San Diego, CA, USA), ANO5 (Abcam, Cambridge, UK), ANO6 and ANO7 (OriGene, Rockville, MD, USA), ANO9 and ANO10 (LSBio, Seattle, WA, USA), and GAPDH (Novus Biologicals, Littleton, CO, USA). Secondary antibodies for automated Western were obtained from Protein Simple (San Jose, CA, USA) or Novus Biologicals (Novus Biologicals, Littleton, CO, USA).

Leitzke S., Seidel J., Ahrens B., Schreiber R., Kunzelmann K., Sperrhacke M., Bhakdi S, & Reiss K. (2022). Influence of Anoctamin-4 and -9 on ADAM10 and ADAM17 Sheddase Function. Membranes, 12(2), 123.

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Measuring Heterogeneity in Platelet Activation

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To better understand the heterogeneity in platelet timing, we measured the time at which single platelets in a 3D clotting environment exposed phosphatidylserine on their outer surface. Phosphatidylserine is a commonly used marker of platelet activation and more broadly of cellular apoptosis. Starting from isolated platelet rich plasma, live platelets were stained with a 1μM Calcien AM (ThermoFisher, L3224) at room temperature for 15 mins. Phosphatidylserine was measured with Annexin V-AF568 (1:20) (ThermoFisher, A13202) and 3d image stacks were collected using a Perkin Elmer UltraVIEW VoX spinning disk confocal with a 20× / 0.75NA objective.
The percentage of PS positive platelets was measured using Volocity 3D imaging software. Platelets stained with calcien were counted at the beginning and end of the experiment with the Volocity built in function, “find objects”, which allowed for consistent tracking of platelet count ensuring experimental platelet count did not change over the course of the experiment. The “find objects” feature was then used to track PS exposed platelets stained with Annexin V-AF568 at each time point. PS exposure data was reported as the percentage of platelets that were PS exposed.

Sun Y., Myers D.R., Nikolov S.V., Oshinowo O., Baek J., Bowie S.M., Lambert T.P., Woods E., Sakurai Y., Lam W.A, & Alexeev A. (2021). Platelet heterogeneity enhances blood clot volumetric contraction: an example of asynchrono-mechanical amplification. Biomaterials, 274, 120828.

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