Apoe3

Manufactured by R&D Systems

APOE3 is a recombinant human Apolipoprotein E3 protein produced in a mouse cell line. Apolipoprotein E is a key player in lipid metabolism and transport. APOE3 is the most common isoform of apolipoprotein E.

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Lab products found in correlation

In situ cell death detection kit, by Roche (1 mentions) Megafasl, by Adipogen (1 mentions) Facsaria, by BD (1 mentions) Facscanto, by BD (1 mentions)

2 protocols using apoe3

Isolation and Co-culture of Murine Immune Cells

Cited 1 time

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Resident and thioglycollate-elicited peritoneal cells, peritoneal macrophages, bone marrow-derived monocytes, brain microglial cell, and photoreceptor outer segment (POS) isolation, and MP-retinal explant co-cultures (all in serum-free X-VIVO 15 medium) were performed as previously described (Sennlaub et al, 2013 (link)). In specific experiments, cells were stimulated with recombinant human CX3CL1 or APOE3 (5 μg/ml, Leinco Technologies), APOE3 (5 μg/ml) with polymyxin B (25 μg/ml, Calbiochem), heat-denatured APOE3 (5 μg/ml, 95°C, 90 min), rat anti-IgG isotype control (100 μg/ml, R&D), rat anti-mouse CD14 (100 μg/ml, R&D), mouse anti-mouse TLR2 (100 μg/ml, Invivogen), and POS prepared as previously described (Molday et al, 1987 (link)). For in vitro apoptosis experiments, 100,000 Mos or Mϕs of the different genotypes were cultured for 24 h with or without MegaFasL (1 ng/ml, AdipoGen). TUNEL staining (In Situ Cell Death Detection Kit, Roche Diagnostics) was performed according to the manufacturer's instructions; TUNEL⁺ and Hoechst⁺ nuclei were counted automatically using the Array Scan (Thermo Fischer).

Levy O., Calippe B., Lavalette S., Hu S.J., Raoul W., Dominguez E., Housset M., Paques M., Sahel J.A., Bemelmans A.P., Combadiere C., Guillonneau X, & Sennlaub F. (2015). Apolipoprotein E promotes subretinal mononuclear phagocyte survival and chronic inflammation in age-related macular degeneration. EMBO Molecular Medicine, 7(2), 211-226.

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Lipoplex and LNP Transfection Assay

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HeLa or HeLa-d1-eGFP cells were plated 24 h before start of incubation. For experiments with a fixed ratio of siRNA to cationic lipid, a 1,000-nM lipoplex solution was mixed (100:4:96, pmol:µl:µl, siRNA:Lipofectamine 2000:OptiMEM). Aliquots of this solution were added to cells in fresh DMEM, with 10% FCS, to obtain the desired final siRNA concentration. After incubation, cells were washed once with PBS, followed by a wash in CellScrub Wash Buffer (Genlantis) and one more PBS wash. Cells were then removed from plates by trypsin digestion, resuspended and washed twice with FACS buffer (PBS, 0.5% BSA, 10 mM HEPES). Fluorescence was analyzed on a FACSCanto (BD) or FACSAria (BD) cell analyzer using FlowJo software (Tree Star).
For LNP incubations, cells were incubated with LNPs dissolved in DMEM, 10% FCS and 1.5 µg/ml recombinant apolipoprotein E3 (ApoE3, R&D Systems). After incubation, cells were washed once with PBS, trypsinized and analyzed by flow cytometry as described above.

Wittrup A., Ai A., Liu X., Hamar P., Trifonova R., Charisse K., Manoharan M., Kirchhausen T, & Lieberman J. (2015). Visualizing lipid-formulated siRNA release from endosomes and target gene knockdown. Nature biotechnology, 33(8), 870-876.

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