Journal: The Journal of biological chemistry

Article Title: A minimally lipidated form of cell-derived apolipoprotein E exhibits isoform-specific stimulation of neurite outgrowth in the absence of exogenous lipids or lipoproteins.

doi: 10.1074/jbc.273.7.4206

Figure Lengend Snippet: FIG. 2. Western blotting of apoE secreted by stably transfected cell lines. Conditioned media (100 mg of protein) were separated by SDS-10% polyacrylamide gel electrophoresis and were electroblotted to a nitrocellulose membrane. The membrane was probed with an affinity purified polyclonal goat anti-human apoE antibody. After incubation with a horseradish peroxidase-conjugated anti-goat IgG, bands were visualized by enhanced chemiluminescence. The slight difference in mobility of the apoE3 and apoE4 bands is due to curvature in the running front and was not seen in other experiments.

Article Snippet: The cells were washed three times with PBS containing 0.5% Triton X-100 and incubated with a rhodamine-conjugated donkey anti-goat IgG antibody (Jackson Immuno Research) at 1:2000 in in PBS containing 1% FBS and 0.5% Triton X-100 for 1 h at 37 °C.

Techniques: Western Blot, Stable Transfection, Transfection, Polyacrylamide Gel Electrophoresis, Membrane, Affinity Purification, Incubation

Journal: The Journal of biological chemistry

Article Title: A minimally lipidated form of cell-derived apolipoprotein E exhibits isoform-specific stimulation of neurite outgrowth in the absence of exogenous lipids or lipoproteins.

doi: 10.1074/jbc.273.7.4206

Figure Lengend Snippet: FIG. 3. Immunocytochemical detection of apoE in stably transfected cell lines. Neuro-2a parental cells (A and B), Neuro-2a apoE3- secreting cells (C), and Neuro-2a apoE4-secreting cells (D) were grown on glass coverslips. ApoE localization was detected with an affinity purified polyclonal goat anti-human apoE antibody and a rhodamine-conjugated secondary antibody (A, C, and D). The white arrows highlight the concentrated localization of apoE in the growth cone domains in the apoE isoform-expressing Neuro-2a cells. B is the transmitted light image of the parental Neuro-2a cells found in A (black arrows highlight 2 Neuro-2a cells). All images were collected by confocal microscopy with a 40 3 oil immersion lens.

Article Snippet: The cells were washed three times with PBS containing 0.5% Triton X-100 and incubated with a rhodamine-conjugated donkey anti-goat IgG antibody (Jackson Immuno Research) at 1:2000 in in PBS containing 1% FBS and 0.5% Triton X-100 for 1 h at 37 °C.

Techniques: Stable Transfection, Transfection, Affinity Purification, Expressing, Confocal Microscopy