Anti-GFP antibody [LGB-1] (ab291)

  Immunocytochemistry/ Immunofluorescence - GFP antibody [LGB-1] (ab291)

Paraformaldehyde fixed COS-7 cells expressing Myr-N15-PAK2-EGFP construct (Vilas et al.(2006) PNAS 103, 6542). Myr-N15-PAK2-EGFP fluorescence is shown in green. Indirect immunofluorescenct detection of N15-PAK-EGFP using ab291 monoclonal LGB-1 anti-GFP at 0.05 ug/ml with chicken anti-mouse secondary antibody conjugated to Alexa594 diluted 1/500 is shown in red. Myr-N15-PAK2-EGFP is localized to membrane ruffles and perinuclear vesicular structures (likely Golgi,TGN or late endosomes).

  Western blot - GFP antibody [LGB-1] (ab291)

All lanes : Anti-GFP antibody [LGB-1] (ab291) at 0.5 µg/ml

Lane 1 : 5ng GFP
Lane 2 : 10ng GFP
Lane 3 : 25ng GFP

Secondary
Sheep anti-mouse IgG HRP conjugate at 1/5000 dilution

Predicted band size : 27 kDa
Observed band size : 27 kDa

  Immunoprecipitation - GFP antibody [LGB-1] (ab291)

ab291 at 6.7µg/mg lysate.
HEK293 Cell lysate at 300µg.
Transfected with CFP-fused protein XXX in pECFP vector. Immunoprecipitation step using Protein G.

This image was kindly supplied by Dr Lindsay Tulloch by Abreview

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  Immunocytochemistry/ Immunofluorescence - GFP antibody [LGB-1] (ab291)

ab291 staining GFP in Dog MDCKII cells transfected with GFP by ICC/IF (Immunocytochemistry/immunofluorescence). Cells were fixed with paraformaldehyde, permeabilized with 0.5% TX100 and blocked with 5% serum for 20 minutes. Samples were incubated with primary antibody (1/250 PBS + 0.1% TX100 + 1% goat serum) for 16 hours at 4°C. An Alexa Fluor^®546-conjugated Goat anti-mouse IgG polyclonal (1/500) was used as the secondary antibody. DAPI was used to stain nuclei. ab291 was used to assess electoporation efficiency of double transfected MDCKII cells.

This image is courtesy of an Abreview submitted by Vladimir Milenkovic

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