rabbit anti apoer  (R&D Systems)

Journal: PLoS ONE

Article Title: Uptake Mechanism of ApoE-Modified Nanoparticles on Brain Capillary Endothelial Cells as a Blood-Brain Barrier Model

doi: 10.1371/journal.pone.0032568

Figure Lengend Snippet: bEnd3 cells were incubated with ApoE-modified nanoparticles (NP-ApoE) or control nanoparticles without ApoE modification (NP-PEG) for 4 h at 37°C and 4°C, respectively. Flow cytometry analysis was performed to quantify their cellular binding. The data are shown as histograms of the FL1-H-channel (autofluorescence of the nanoparticles) as well as in the table with the analysis of the Y mean fluorescence and the percentage of positive cells. Green: NP-ApoE, red: NP-PEG, blue: untreated control.

Article Snippet: Then, the cells were permeabilized, fixed and blocked with 5% goat serum (20 min, 4°C) before they were incubated with the primary antibodies against the ApoE receptor (ApoER) (Acris Antibodies GmbH, Herford, Germany), Megalin (abcam, Cambridge, UK), LDLR (abcam, Cambridge, UK) or LRP1 (Pietrzik et al. 2002 ) for 30 min at 4°C.

Techniques: Incubation, Modification, Control, Flow Cytometry, Binding Assay, Fluorescence

Journal: PLoS ONE

Article Title: Uptake Mechanism of ApoE-Modified Nanoparticles on Brain Capillary Endothelial Cells as a Blood-Brain Barrier Model

doi: 10.1371/journal.pone.0032568

Figure Lengend Snippet: The green autofluorescence of the nanoparticles was used for detection. The cytosol was stained in red with CellTracker™ Red CMTPX, and the nucleus was stained in blue with DAPI. Pictures were taken within inner sections of the cells. Untreated control cells: a) overlay of all fluorescence channels, b) display of the blue nucleus channel, c) display of the green nanoparticle channel, d) display of the red cytosol channel. Cells with the unspecific control NP-PEG: e) overlay of all fluorescence channels, f) display of the blue nucleus channel, g) display of the green nanoparticle channel, h) display of the red cytosol channel. Cells with the specific NP-ApoE: i) overlay of all fluorescence channels, j) display of the blue nucleus channel, k) display of the green nanoparticle channel, l) display of the red cytosol channel.

Article Snippet: Then, the cells were permeabilized, fixed and blocked with 5% goat serum (20 min, 4°C) before they were incubated with the primary antibodies against the ApoE receptor (ApoER) (Acris Antibodies GmbH, Herford, Germany), Megalin (abcam, Cambridge, UK), LDLR (abcam, Cambridge, UK) or LRP1 (Pietrzik et al. 2002 ) for 30 min at 4°C.

Techniques: Staining, Control, Fluorescence

Journal: PLoS ONE

Article Title: Uptake Mechanism of ApoE-Modified Nanoparticles on Brain Capillary Endothelial Cells as a Blood-Brain Barrier Model

doi: 10.1371/journal.pone.0032568

Figure Lengend Snippet: Determination of the receptor state of the bEnd3 cells.

Article Snippet: Then, the cells were permeabilized, fixed and blocked with 5% goat serum (20 min, 4°C) before they were incubated with the primary antibodies against the ApoE receptor (ApoER) (Acris Antibodies GmbH, Herford, Germany), Megalin (abcam, Cambridge, UK), LDLR (abcam, Cambridge, UK) or LRP1 (Pietrzik et al. 2002 ) for 30 min at 4°C.

Techniques:

Journal: PLoS ONE

Article Title: Uptake Mechanism of ApoE-Modified Nanoparticles on Brain Capillary Endothelial Cells as a Blood-Brain Barrier Model

doi: 10.1371/journal.pone.0032568

Figure Lengend Snippet: Co-incubation of bEnd3 cells with the different nanoparticulate formulations and LDL.

Article Snippet: Then, the cells were permeabilized, fixed and blocked with 5% goat serum (20 min, 4°C) before they were incubated with the primary antibodies against the ApoE receptor (ApoER) (Acris Antibodies GmbH, Herford, Germany), Megalin (abcam, Cambridge, UK), LDLR (abcam, Cambridge, UK) or LRP1 (Pietrzik et al. 2002 ) for 30 min at 4°C.

Techniques: Control

Journal: PLoS ONE

Article Title: Uptake Mechanism of ApoE-Modified Nanoparticles on Brain Capillary Endothelial Cells as a Blood-Brain Barrier Model

doi: 10.1371/journal.pone.0032568

Figure Lengend Snippet: Co-incubation of bEnd3 cells with the different nanoparticulate formulations and LDL+RAP.

Article Snippet: Then, the cells were permeabilized, fixed and blocked with 5% goat serum (20 min, 4°C) before they were incubated with the primary antibodies against the ApoE receptor (ApoER) (Acris Antibodies GmbH, Herford, Germany), Megalin (abcam, Cambridge, UK), LDLR (abcam, Cambridge, UK) or LRP1 (Pietrzik et al. 2002 ) for 30 min at 4°C.

Techniques: Control

Journal: PLoS ONE

Article Title: Uptake Mechanism of ApoE-Modified Nanoparticles on Brain Capillary Endothelial Cells as a Blood-Brain Barrier Model

doi: 10.1371/journal.pone.0032568

Figure Lengend Snippet: Co-incubation of bEnd3 cells with the different nanoparticulate formulations and LRP1 Dom II and LRP1 Dom IV, respectively.

Article Snippet: Then, the cells were permeabilized, fixed and blocked with 5% goat serum (20 min, 4°C) before they were incubated with the primary antibodies against the ApoE receptor (ApoER) (Acris Antibodies GmbH, Herford, Germany), Megalin (abcam, Cambridge, UK), LDLR (abcam, Cambridge, UK) or LRP1 (Pietrzik et al. 2002 ) for 30 min at 4°C.

Techniques: