GFP antibody

Catalog No. ABIN294468

Product Details anti-GFP Antibody

Target: GFP (Green Fluorescent Protein (GFP))

Reactivity: Aequorea victoria

Host: Chicken

Clonality: Polyclonal

Conjugate: This GFP antibody is un-conjugated

Application: Western Blotting (WB), Immunoprecipitation (IP), Immunocytochemistry (ICC)

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

Purification: Purified

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

Isotype: IgY

Application Details

Application Notes: Approved: ICC, IP, WB

Usage: 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 μg 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.

Comment:

Target Species of Antibody: Aequorea victoria

Restrictions: For Research Use only

Handling

Format: Liquid

Concentration: Lot specific

Buffer: 10 mM sodium chloride, 0.02 % sodium azide.

Preservative: Sodium azide

Precaution of Use: This product contains Sodium azide: a POISONOUS AND HAZARDOUS SUBSTANCE which should be handled by trained staff only.

Handling Advice: Avoid repeat freeze-thaw cycles.

Storage: 4 °C,-20 °C

Storage Comment: Short term: 4°C. Long term: Store at -20°C. Avoid freeze-thaw cycles.

Target Details for GFP

Target: GFP (Green Fluorescent Protein (GFP))

Alternative Name: GFP (GFP Products)

Synonyms: green fluorescent protein antibody, gfp antibody

Background: Name/Gene ID: GFP

Synonyms: GFP

UniProt: P42212