Journal: eLife

Article Title: Membrane characteristics tune activities of endosomal and autophagic human VPS34 complexes

doi: 10.7554/eLife.58281

Figure Lengend Snippet:

Article Snippet: Chemical compound, drug , DSPE-PEG(2000) Biotin , Avanti Polar Lipids, Inc , 880129C , Dissolved in chloroform.

Techniques: Recombinant

Journal: Nanoscale

Article Title: Incorporating Functionalized Polyethylene Glycol Lipids into Reprecipitated Conjugated Polymer Nanoparticles for Bioconjugation and Targeted Labeling of Cells

doi: 10.1039/c0nr00746c

Figure Lengend Snippet: Size of PFBT nanoparticles prepared with different varieties of PEG lipids by TEM.

Article Snippet: Streptavidin Pull-down of Biotin PEG Lipid-PFBT nanoparticles PFBT nanoparticles (28 ppm) prepared with biotin functionalized PEG lipid were incubated with magnetic streptavidin beads (New England Biolabs) for 30 mins in phosphate buffered saline (PBS; 140 mM NaCl, 2.7 mM KCl, 10 mM Na 2 HPO 4 , 1.8 mM KH 2 PO 4 ).

Techniques:

Journal: Nanoscale

Article Title: Incorporating Functionalized Polyethylene Glycol Lipids into Reprecipitated Conjugated Polymer Nanoparticles for Bioconjugation and Targeted Labeling of Cells

doi: 10.1039/c0nr00746c

Figure Lengend Snippet: (A) TEM image of methoxy 2000 Mr PEG lipid-PFBT nanoparticles. Images were acquired at 120 kV on a cryostage at liquid nitrogen temperature and are shown at 120,000 magnification. Scale bar is 500 nm. Some blurring of the borders between these small nanoparticles is an artefact of high magnification. (B) Histogram of size distribution and fit to a Gaussian function for the methoxy 2000 Mr PEG lipid-PFBT nanoparticles showing the average diameter to be 24 ± 5 nm (mean ± FWHM).

Article Snippet: Streptavidin Pull-down of Biotin PEG Lipid-PFBT nanoparticles PFBT nanoparticles (28 ppm) prepared with biotin functionalized PEG lipid were incubated with magnetic streptavidin beads (New England Biolabs) for 30 mins in phosphate buffered saline (PBS; 140 mM NaCl, 2.7 mM KCl, 10 mM Na 2 HPO 4 , 1.8 mM KH 2 PO 4 ).

Techniques:

Journal: Nanoscale

Article Title: Incorporating Functionalized Polyethylene Glycol Lipids into Reprecipitated Conjugated Polymer Nanoparticles for Bioconjugation and Targeted Labeling of Cells

doi: 10.1039/c0nr00746c

Figure Lengend Snippet: Quantum yields for PFBT CPNs (fluorescein in 0.1 M NaOH as reference).

Article Snippet: Streptavidin Pull-down of Biotin PEG Lipid-PFBT nanoparticles PFBT nanoparticles (28 ppm) prepared with biotin functionalized PEG lipid were incubated with magnetic streptavidin beads (New England Biolabs) for 30 mins in phosphate buffered saline (PBS; 140 mM NaCl, 2.7 mM KCl, 10 mM Na 2 HPO 4 , 1.8 mM KH 2 PO 4 ).

Techniques:

Journal: Nanoscale

Article Title: Incorporating Functionalized Polyethylene Glycol Lipids into Reprecipitated Conjugated Polymer Nanoparticles for Bioconjugation and Targeted Labeling of Cells

doi: 10.1039/c0nr00746c

Figure Lengend Snippet: Apparent hydrodynamic size for carboxy 2000 M r PEG lipid-PFBT CPNs prepared with a range of starting PEG lipid concentrations

Article Snippet: Streptavidin Pull-down of Biotin PEG Lipid-PFBT nanoparticles PFBT nanoparticles (28 ppm) prepared with biotin functionalized PEG lipid were incubated with magnetic streptavidin beads (New England Biolabs) for 30 mins in phosphate buffered saline (PBS; 140 mM NaCl, 2.7 mM KCl, 10 mM Na 2 HPO 4 , 1.8 mM KH 2 PO 4 ).

Techniques:

Journal: Nanoscale

Article Title: Incorporating Functionalized Polyethylene Glycol Lipids into Reprecipitated Conjugated Polymer Nanoparticles for Bioconjugation and Targeted Labeling of Cells

doi: 10.1039/c0nr00746c

Figure Lengend Snippet: Streptavidin coated magnetic beads were incubated with biotinylated 2000 Mr PEG lipid PFBT nanoparticles, washed to remove unbound CPNs, and then incubated with free biotin to release bound nanoparticles. Magnetic beads were removed with a strong magnet, and the fluorescence of the supernatant was recorded (solid line; λex = 460 nm ). Control experiments using carboxy 2000 Mr PEG lipid-PFBT nanoparticles do not bind streptavidin magnetic beads, as shown by the emission spectrum of the control supernatant (dashed line).

Article Snippet: Streptavidin Pull-down of Biotin PEG Lipid-PFBT nanoparticles PFBT nanoparticles (28 ppm) prepared with biotin functionalized PEG lipid were incubated with magnetic streptavidin beads (New England Biolabs) for 30 mins in phosphate buffered saline (PBS; 140 mM NaCl, 2.7 mM KCl, 10 mM Na 2 HPO 4 , 1.8 mM KH 2 PO 4 ).

Techniques: Magnetic Beads, Incubation, Fluorescence

Journal: Nanoscale

Article Title: Incorporating Functionalized Polyethylene Glycol Lipids into Reprecipitated Conjugated Polymer Nanoparticles for Bioconjugation and Targeted Labeling of Cells

doi: 10.1039/c0nr00746c

Figure Lengend Snippet: (A) Representative fluorescence image of streptavidin-coated cover glass incubated with biotinylated 2000 Mr PEG-lipid-PFBT nanoparticles (λex = 495 nm; λem = 510 nm long pass filter). Scale bar = 10 μm. (B) histogram of biotinylated 2000 Mr PEG-lipid-PFBT nanoparticle fluorescence intensities obtained from the image; a threshold mask was applied to all objects(C) fluorescence image of streptavidin-coated glass slide incubated with carboxy 2000 Mr PEG lipid-PFBT nanoparticles, as a control for nonspecific binding to slide and (D) histogram of the 2000 Mr PEG lipid-PFBT nanoparticle fluorescence intensities obtained from the image. Exposure times were identical for (A) and (C). The diffraction limit of the microscope was 225 nm.

Article Snippet: Streptavidin Pull-down of Biotin PEG Lipid-PFBT nanoparticles PFBT nanoparticles (28 ppm) prepared with biotin functionalized PEG lipid were incubated with magnetic streptavidin beads (New England Biolabs) for 30 mins in phosphate buffered saline (PBS; 140 mM NaCl, 2.7 mM KCl, 10 mM Na 2 HPO 4 , 1.8 mM KH 2 PO 4 ).

Techniques: Fluorescence, Incubation, Binding Assay, Microscopy

Journal: Nanoscale

Article Title: Incorporating Functionalized Polyethylene Glycol Lipids into Reprecipitated Conjugated Polymer Nanoparticles for Bioconjugation and Targeted Labeling of Cells

doi: 10.1039/c0nr00746c

Figure Lengend Snippet: (A) Differential Interference Image (DIC) of fixed J774A.1 cells; (B) fluorescence image of fixed J774A.1 cells labeled with biotinylated PFBT nanoparticles. Scale bar is 50μm. J774 A1 macrophage cells which express CD16/32 (Fc receptor) were paraformaldehyde fixed and incubated with biotinylated anti-CD16/32 antibody. After washing the cell with RB, the Cells were next incubated with streptavidin, washed, and labeled with biotinylated PEG lipid-PFBT nanoparticles. Images were obtained with 495 nm excitation, using a 510 nm long pass emission filter.

Article Snippet: Streptavidin Pull-down of Biotin PEG Lipid-PFBT nanoparticles PFBT nanoparticles (28 ppm) prepared with biotin functionalized PEG lipid were incubated with magnetic streptavidin beads (New England Biolabs) for 30 mins in phosphate buffered saline (PBS; 140 mM NaCl, 2.7 mM KCl, 10 mM Na 2 HPO 4 , 1.8 mM KH 2 PO 4 ).

Techniques: Fluorescence, Labeling, Incubation

Journal: bioRxiv

Article Title: Characterization of gustatory receptor 7 in the brown planthopper reveals functional versatility

doi: 10.1101/2020.09.05.284208

Figure Lengend Snippet: Nl Gr7 recombinant protein and Nl Gr7-ligands overlay assays. (A) Purified Nl Gr7, (B) Nl Gr7-lectin overlay assay shows that Nl Gr7 binds to several lectins. Protein-lipid overlay assay shows that Nl Gr7 binds to several anionic lipids. (C) Nl Gr7 reveals interaction to phospholipids, and (D) interaction to sphingolipids.

Article Snippet: Additional biotinylated lectins (20 µg/ 50 µl; Vector Labs, Burlingame, CA) were added to each well and were incubated for 2 h at room temperature.

Techniques: Recombinant, Purification, Overlay Assay, Protein-lipid Overlay Assay (PLOA)