AnaSpec is pleased to expand our GFP antibodies offerings. The new GFP antibodies include two monoclonals raised in mouse and a third, a polyclonal raised in chicken. Purfied green-fluorescent protein (GFP) was used as antigen. Antibody reactivity was confirmed by ELISA. Specificity was confirmed by western blot using GFP protein. These antibodies also recognize other GFP mutant forms.

Green fluorescent protein (GFP) is a 27-kDa protein which was originally cloned from Jellyfish cnidarians, Aequorea victoria. This exceptional protein absorbs blue light (maximally at 395 nm) and emits green light (peak at 509 nm) without any requirement for exogenous substrates and cofactors (1). These unique intrinsic fluorescence properties make GFP an invaluable tool in cell biology research in applications such as monitoring gene expression and protein localization of GFP-tagged proteins in vivo. Other applications of GFP include assessment of protein-protein interactions through the yeast two hybrid system and measurement of distance between proteins through fluorescence energy transfer (FRET) protocols. Several mutant forms of GFP have been developed which fluoresce more intensely and have shifted excitation maximum when compared to the wild type GFP, making them useful for FACS, fluorescence microscopy, and double-labeling applications (2, 3).

		Figure 1. GFP protein (~150 ng) was probed on the Western blot using anti-GFP-Tag chicken IgY antibody (1:1,000 dilution)(Cat.# 55423). Secondary rabbit anti-chicken IgY-HilyteFluor™ 680 nm labeled antibody (Cat.# 29709-H680) was used to visualize fluorescent band in the near-infrared part of the light spectra. An immunoreactive band was detected at ~27 kDa.
		Figure 2. Recombinant GFP protein (5 mg) was probed on the Western Blot using anti-GFP (1A5) mouse monoclonal antibody (Cat# 55887).  Secondary goat anti-mouse IgG-HiLyte FlourTM 750 nm labeled antibody (Cat# 61057-H750) was used at a 1/25,000 dilution to visualize fluorescent band in the near-infrared part of the light spectra.  An immunoreactive band at 27 kDa was detected.
		Figure 3. IP: GFP protein (15 mg) was immunoprecipitated with the mouse anti-GFP Tag monoclonal antibody (Cat# 55886) and probed with a rabbit anti-GFP Tag polyclonal antibody (Cat#29779) at 1 mg/ml.
		Figure 4. HEK293 cells transfected with GFP were probed with mAb Anti-GFP clone 6A2B3 (Cat# 55886) and detected with goat anti-mouse IgG conjugated with HiLyte Fluor™ 555 secondary antibody (Cat# 61057-05-H555). Green color shows GFP and yellow color is a combination of the green (GFP) and red (HiLyte Fluor™ 555).

Click here for a full listing of other Tag Antibodies, available as fluorogenic labeled, chromogenic labeled or unlabeled.

References

1. Cormack, B. et al. Gene 173, 33 (1996).
2. Rizzuto, R. et al. Curr Biol 6, 183 (1996).
3. Chalfie, M. et al. Science 263, 802 (1994).