Application DetailsWestern blotting (1:1,000-1:5,000) and Immunocytochemistry (1:2,000-1:5,000). Biosensis recommends optimal dilutions/concentrations should be determined by the end user.
Product ValidationAntibody recognizes GFP protein in GFP-transfected HEK293 cells, but not in non-transfected control cells.
TargetGreen fluorescent protein (GFP)
SpecificitySpecific for GFP, does not cross-react with mCherry.
Target Host SpeciesJellyfish
Species ReactivitySpecies Independent
Antibody HostGoat
Antibody TypePolyclonal
Antibody IsotypeIgG
ConjugateUnconjugated
Immunogen DescriptionRecombinant AcGFP protein expressed in and purified from E. Coli.
Immunogen LengthFull-length recombinant protein.
Positive ControlGFP-transfected HEK293 cells.
Negative ControlNon-transfected HEK293 cells.
Purity DescriptionAffinity-purified from goat serum using the immunogen.
Physical StateSolid.
FormatLyophilized from PBS buffer pH 7.2-7.6 with 0.1% trehalose, without preservatives
Reconstitution InstructionsSpin vial briefly before opening. Reconstitute in 100 µL sterile-filtered, ultrapure water. Centrifuge to remove any insoluble material.
Storage InstructionsStore lyophilized antibody at 2-8°C. After reconstitution divide into aliquots and store at -20°C for long-term storage. Store at 2-8°C short-term (up to 4 weeks). Avoid repetitive freeze/thaw cycles.
Batch NumberPlease see item label.
Expiration Date12 months after date of receipt (unopened vial).
Scientific BackgroundThe green fluorescent protein (GFP) is a 27 kDa protein isolated originally from the jellyfish Aequoria victoria. It has an endogenous fluorochrome activity with excitation maximum at 395 nm and emission maximum at 509 nm, which is similar to that of fluorescein. GFP can be expressed in fluorescent form in essentially any prokaryotic or eukaryotic cell.
This GFP rabbit antibody was made against a recombinant GFP construct originating from an Aequoria species which was engineered to improve spectral properties and prevent oligomerization. This form of GFP, referred to as AcGFP, is 94% identical to the eGFP developed by Tsien and co-workers. The antibody can be used to verify the expression, size and stability of both AcGFP and eGFP fusion proteins in western blotting experiments and to amplify GFP signals in tissues of transgenic animals.
Left: Analysis of GFP fusion protein-transfected HEK293 cells by Immunocytochemistry. GFP expression (green) was detected with goat antibody to GFP (red, 1:5,000). Blue: Hoechst nuclear stain. The goat antibody reveals GFP protein expressed only in transfected cells, and as a result these cells appear yellow in color. Top: merged image; bottom panel: individual fluorescence channels. Right: Western blot analysis of GFP expression (green) in transfected HEK293 cells with goat anti-GFP antibody (1:1,000). Lane 1: molecular weight marker; Lane 2: non-transfected control cells; Lane 3: GFP-transfected cells; Lane 4: mCherry-transfected cells. A strong band at ~27 kDa corresponds to GFP protein detected only in cells transfected with GFP construct. This antibody does not recognize the mCherry protein.