For western blot analysis, whole cell extracts were prepared from approximately 5×106 cells by trichloroacetic acid (TCA) precipitation as described previously [29 (link)]. To detect GFP-fused proteins, a rabbit polyclonal anti-GFP antibody (600-401-215, Rockland Immunochemicals Inc.) was used. To detect spNup98, a mouse monoclonal anti-Nup98 antibody (13C2) was used [45 (link)]. HRP-conjugated goat anti-rabbit or anti-mouse IgG (NA9340-1ml or NA9310-1ml, GE Healthcare) was used as a secondary antibody. Protein bands were detected by chemiluminescence using ChemiDoc MP imaging system (Bio-Rad).
Anti gfp antibody 600 401 215
Manufactured by Rockland Immunochemicals
The Anti-GFP antibody (600-401-215) is a laboratory reagent used to detect the presence of green fluorescent protein (GFP) in samples. It is a polyclonal antibody that specifically binds to GFP, enabling researchers to identify and quantify GFP-tagged proteins or cells.
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2 protocols using anti gfp antibody 600 401 215
1
Western Blot Analysis of GFP-Nup98 Proteins
2
Immunohistochemical Staining of GFP
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For immunohistochemical and immunofluorescence studies, 5 um thick bundled paraffin-embedded wax tissue block sections were used. After rehydrating the sections in xylene and alcohol, they were treated with an antigen unmasking solution (H-3300, Vector
Laboratories) and microwaved. The endogenous peroxidase activity was quenched using 0.3% hydrogen peroxide (H/1750/17, Fisher Scientific) solution in methanol, and incubated for 30 minutes in a histology dish and washed in PBS. Sections were incubated with 1:500 anti-GFP antibody (600-401-215, Rockland) after blocking non-specific binding. 100 l of Dako envision goat anti-rabbit secondary antibody (K4002, Dako) was used as secondary antibody before the sections were incubated in the peroxidase substrate (SK-4100, Vector
Laboratories) and microwaved. The endogenous peroxidase activity was quenched using 0.3% hydrogen peroxide (H/1750/17, Fisher Scientific) solution in methanol, and incubated for 30 minutes in a histology dish and washed in PBS. Sections were incubated with 1:500 anti-GFP antibody (600-401-215, Rockland) after blocking non-specific binding. 100 l of Dako envision goat anti-rabbit secondary antibody (K4002, Dako) was used as secondary antibody before the sections were incubated in the peroxidase substrate (SK-4100, Vector
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