Anti-GFP 抗体 [LGB-1]

• ICC/IF

Lab

Immunocytochemistry/ Immunofluorescence - Anti-GFP antibody [LGB-1] (AB291)

ab291 staining GFP in GFP-transfected NIH3T3 cells. The cells were fixed with 4% formaldehyde (10min) and then blocked in 1% BSA / 0.3M glycine in 0.1%PBS-Tween for 1h. The cells were then incubated with ab291 at 1/200 dilution overnight at +4°C followed by incubation with ab150117, Goat Anti-Mouse IgG H&L (Alexa Fluor® 488), for 1 hour, at 1μg/ml.
Under identical experimental conditions, when compared to the basal level of GFP expression in transfected NIH3T3 cells, the cells upon which ab291 was applied gave a stronger signal in the 488 channel, indicating that ab291 is binding to GFP and therefore eliciting signal amplification.
ab291 was also applied to non-GFP-transfected NIH3T3 cells, which produced no positive staining, indicating specificity for GFP. Nuclear DNA was labelled with 1.43μM DAPI (blue).

• IP

AbReview16870****

Immunoprecipitation - Anti-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.

All lanes:

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

Predicted band size: 27 kDa

false

This image was kindly supplied by Dr Lindsay Tulloch by Abreview

• ICC/IF

AbReview19325****

Immunocytochemistry/ Immunofluorescence - Anti-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

• ICC/IF

Unknown

Immunocytochemistry/ Immunofluorescence - Anti-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).

• WB

Unknown

Western blot - Anti-GFP antibody [LGB-1] (AB291)

All lanes:

Western blot - Anti-GFP antibody [LGB-1] (ab291) at 0.5 µg/mL

Lane 1:

5ng GFP

Lane 2:

10ng GFP

Lane 3:

25ng GFP

Secondary

All lanes:

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

Predicted band size: 27 kDa

Observed band size: 27 kDa

false