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You are here:Home » Antibodies » Abs to Assay Detection » Anti -Green Fluorescent Protein (GFP)

Anti -Green Fluorescent Protein (GFP)

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Specifications

Clone Host Grade Applications
Polyclonal Chicken Purified B IP IC
Catalog #G8965-09
ApplicationsSuitable for use in Western Blot, Immunocytochemistry and Immunoprecipitation. Other applications have not been tested.
Recommended DilutionsWestern 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 4°C, 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 4°C for short-term only. Aliquot to avoid repeated freezing and thawing. Store at -20°C. 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.
Clone TypePolyclonal
IsotypeIgY
HostChicken
Concentration~1mg/ml
FormSupplied as a liquid in TBS, pH 7.5, <0.05% sodium azide.
PurityPurified
ImmunogenAffinity purified, recombinant green fluorescent protein (GFP) containing a 6-His tag (single band, SDS-PAGE).
SpecificityRecognizes green fluorescent protein (GFP). Has been used to detect GFP in primary neurons and zebrafish fibroblasts.
Important NoteThis product as supplied is intended for research use only, not for use in human, therapeutic or diagnostic applications without the expressed written authorization of United States Biological.


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