Technical Data
Green Fluorescent Protein (GFP)
Suitable for use in Western Blot, Immunocytochemistry and Immunoprecipitation. Other applications have not been tested.

Recommended Dilutions:
Western Blot: Detects a band of the proper molecular weight, 30kD, in lysates from E. coli expressing GFP, but not in E. coli that do not express or in lysates prepared from E. coli containing the GFP-expressing plasmid but not induced, detect GFP in lysates from COS or primary neurons or fibroblast cells transfected with plasmids directing expression GFP or a tau-GFP fusion protein. Did not detect any signal when 50ug of protein from a rat brain lysate (negative control) was fractionated on the SDS-PAGE. Titers were 1:5000-1:10,000 when detected with an anti-chicken HRP conjugated secondary by ECL or by a colorimetric assay using DAB.
Immunocytochemistry: GFP can be detected in cells that are transfected with a plasmid directing the expression of either GFP or a GFP-Fusion protein. Cells that do not express GFP exhibit no detectable staining. Binding was determined using HRP conjugated, anti-chicken secondary antibodies and visualized using DAB. The anti-GFP antibodies were used at dilutions of 1:1000-1:2500. Cells were fixed using 4% formaldehyde in PBS, pH 7.4. For immunocytochemistry, the cultures were fixed in 4% formaldehyde in PBS, pH 7.4, for 20 minutes at RT. The cells are permeabilized with 0.2% Triton X-100 and 2% serum (corresponding to the animal in which the secondary antibody was raised) in PBS for 20 minutes at RT. Cell cultures are then incubated with primary antisera in PBS containing 2% serum (as above), at 4C, overnight. Has also been used successfully with the avidin/biotin/peroxidase detection system. Cultures were washed four times (10 minutes each) between steps, and reaction product is developed with a 0.05% diaminobenzidine/ 0.02% hydrogen peroxide solution on ice for 8-10 minutes.
Optimal dilutions to be determined by the researcher.
Storage and Stability:
May be stored at 4C for short-term only. Aliquot to avoid repeated freezing and thawing. Store at -20C. Aliquots are stable for at least 12 months. For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap.
250ug-20CBlue IceChicken
Affinity purified, recombinant green fluorescent protein (GFP) containing a 6-His tag (single band, SDS-PAGE).
Supplied as a liquid in TBS, pH 7.5, <0.05% sodium azide.
Recognizes green fluorescent protein (GFP). Has been used to detect GFP in primary neurons and zebrafish fibroblasts.
Intended for research use only. Not for use in human, therapeutic, or diagnostic applications.
1. Hallam, S.J., et al., Nature Neuroscience 5: 1137-1146 (2002). 2. Sippy, T., et al., Nature Neuroscience 10: 1031-1038 (2003). 3. Jakobsson, J., et al., European Journal of Neuroscience 19: 761-765 (2004). 4. Zhao, S., et al., European Journal of Neuroscience 19: 1133-1140 (2004). 5. Yawata, S., et al., Journal of Neuroscience 26: 3626-3633 (2006).