Aequorea victoria GFP

Chicken

Aequorea victoria (tested or 100% immunogen sequence identity)

IgY Polyclonal

Unconjugated. Also available conjugated with APC, Biotin, FITC, HRP, PE.

Purified

Unmodified

Affinity purified, recombinant green fluorescent protein (GFP) containing a 6-His tag (single band, SDS-PAGE).

Recognizes green fluorescent protein (GFP). Has been used to detect GFP in primary neurons and zebrafish fibroblasts.

• ICC
• Western blot
• Immunoprecipitation

Suitable for use in Western Blot, Immunocytochemistry and Immunoprecipitation. 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 50 ug of protein from a rat brain lysate (negative control) was fractionated on the SDS-PAGE. Titers were 1:5000-1:10000 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. Has been used to detect GFP in primary neurons and zebrafish fibroblasts.

10 mM sodium chloride, 0.02% sodium azide.

Short term: store at 4°C. Long term: store at -20°C. Avoid freeze-thaw cycles.

For research use only. Intended for use by laboratory professionals.

This antibody carries the LSBio 100% Guarantee.

Uniprot: P42212 NCBI: M62653 P42212