Mfg e8 fitc

Manufactured by Prolytix

MFG-E8-FITC is a fluorescently labeled protein that binds to phosphatidylserine. It can be used in flow cytometry, immunohistochemistry, and other applications to detect and quantify cells undergoing apoptosis.

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Lab products found in correlation

Model 963, by Kopf Instruments (1 mentions) Annexin 5 fitc, by Thermo Fisher Scientific (1 mentions) Annexin 5 apc, by Thermo Fisher Scientific (1 mentions) Sytox green, by BD (1 mentions) Sytox green, by Thermo Fisher Scientific (1 mentions) Facscalibur, by BD (1 mentions)

2 protocols using mfg e8 fitc

In Vivo Molecular Labeling of Neural Cells

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Mice were anesthetized with an injection of a mixture of medetomidine/midazolam/butorphanol (MMB; 0.75 mg/kg, 4 mg/kg, and 5 mg/kg, respectively), and the head was immobilized on the stereotaxic stage (David Kopf Instruments; model 963) using ear bars. A 1.5 μl volume of PSVue 480 (1 mM; Molecular Targeting Technologies, Inc.), PSVue 550 (1 mM; Molecular Targeting Technologies, Inc.), AnnexinV-FITC (MBL Co., Ltd.), or MFG-E8-FITC (83 µg/ml; Haematologic Technologies, LLC) was stereotaxically injected into the OB (5.0 mm anterior, 0.9 mm lateral to bregma, and 1.5–2.0 mm deep) or hippocampal DG (1.4 mm posterior, 0.6 mm lateral to bregma, and 1.9–2.4 mm deep) of male adult Dcx-CreER^T2; R26-tdTomato, Dcx-CreER^T2; R26-EPT; R26-tdTomato, Dcx-CreER^T2; R26-D89E; R26-tdTomato, and WT mice as reported previously (Sawada et al., 2018 (link)). These mice were fixed 3 h after injection by transcardiac perfusion with 4% PFA/0.1 M phosphate buffer (PB).

Kurematsu C., Sawada M., Ohmuraya M., Tanaka M., Kuboyama K., Ogino T., Matsumoto M., Oishi H., Inada H., Ishido Y., Sakakibara Y., Nguyen H.B., Thai T.Q., Kohsaka S., Ohno N., Yamada M.K., Asai M., Sokabe M., Nabekura J., Asano K., Tanaka M, & Sawamoto K. (2022). Synaptic pruning of murine adult-born neurons by microglia depends on phosphatidylserine. The Journal of Experimental Medicine, 219(4), e20202304.

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Analyzing Cell Death Mechanisms

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All in vitro experiments assessed by flow cytometry included at least triplicate cultures for each condition (n≥3), and data shown are representative of at least two independent experiments. To assess necroptosis or apoptosis, cells were trypsinized, resuspended in serum-free DMEM and stained with Annexin V-APC (1:50, eBioscience), 50 nM SytoxGreen (invitrogen) or 2 μg/ml PI (Sigma) for 5min. Alternatively, cells were stained with Annexin V-FITC (1:50, eBioscience) or MFG-E8-FITC (1:50, Haematologic Technologies) and TO-PRO^®-3 (500 nM, Molecular Probes).
Cells with Annexin-V^+/−, PI^+/−, TO-PRO^®-3^+/− or SytoxGreen^+/− were analyzed by flow cytometry using FACScan or FACsCalibur systems (BD Biosciences) and FlowJo Collectors’ Edition software (Tree Star). The percentages of differently labeled cells were calculated by FlowJo software (Tree Star).

Gong Y.N., Guy C., Olauson H., Becker J.U., Yang M., Fitzgerald P., Linkermann A, & Green D.R. (2017). ESCRT-III acts downstream of MLKL to regulate necroptotic cell death and its consequences. Cell, 169(2), 286-300.e16.

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