Anti-GFP antibody (ab1218)

  Immunoprecipitation - GFP antibody (ab1218)

Immunoprecipitation using ab1218 at 0.01µl/µl HEK (human embryonic kidney) 293 whole cell lysate incubated with protein G matrix for 1 hour at 4°C.

Total protein input was 3,000,000 cells.

Lane 1: GFP-RhoA wt

Lane 2: GFP-Rac1 wt

Lane 3: GFP-empty

The western blot antibody was ab290 (undiluted).

This image is courtesy of an Abreview submitted by Dr Peter Jordan

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  Immunocytochemistry/ Immunofluorescence - GFP antibody (ab1218)

ab1218 staining GFP in human prostate epithelial primary cells by Immunocytochemistry/ Immunofluorescence. Cells (ARPE-19 cells transfected with mouse Rab27A-GFP) were fixed with paraformaldehyde, permeabilized with 0.5% Triton x100 and blocking with 5% serum was performed for 20 minutes at 25°C. Samples were incubated with primary antibody (1/500: in 1% goat serum, 0.1%TX100, 1XPBS) for 16 hours at 4°C. An Alexa Fluor^®546-conjugated goat polyclonal to mouse IgG was used as secondary antibody at 1/500 dilution. DAPI was used to stain the cell nuclei (blue) and mRab27A-GFP with ab1218 (red).

This image is a courtesy of Vladimir Milenkovic

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  Western blot - GFP antibody (ab1218)

Anti-GFP antibody (ab1218) at 1/1000 dilution + Human HEK293 whole cell lysate at 3 µg

Secondary
A Pacific Blue^TM-conjugated Goat anti-mouse IgG Monoclonal at 1/3000 dilution
developed using the ECL technique

Observed band size : 120 kDa (why is the actual band size different from the predicted?)


Exposure time : 40 seconds

Blocking STep: 5% Milk for 45 minutes at 24°C

GST fusion protein ectopically expressed in HEK293 cells.

This image is courtesy of an anonymous Abreview

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