GFP Antibody

Description

Background

Green fluorescent protein is a 27 kDa protein produced from the jellyfish Aequorea victoria, which emits green light (emission peak at a wavelength of 509nm) when excited by blue light. GFP is an important tool in cell biology research. GFP is widely used enabling researchers to visualize and localize GFP-tagged proteins within living cells without the need for chemical staining.

Application Note

Anti-GFP is designed to detect GFP and its variants. Goat Anti-GFP has been tested by ELISA, SDS-PAGE, Western blot, and Immunofluorescence. This product is ideal for western blotting, ELISA, immunofluorescence, IHC, and IP. This antibody can be used to detect GFP by ELISA (sandwich or capture) for the direct binding of antigen and recognizes wild type, recombinant and enhanced forms of GFP. Biotin conjugated polyclonal anti-GFP used in a sandwich ELISA is well suited to titrate GFP in solution using this antibody in combination with Rockland's monoclonal anti-GFP (600-301-215) using either form of the antibody as the capture or detection antibody. However, use the monoclonal form only for the detection of wild type or recombinant GFP as this form does not sufficiently detect 'enhanced' GFP. The detection antibody is typically conjugated to biotin and subsequently reacted with streptavidin-HRP (code # S000-03). Fluorochrome conjugated polyclonal anti-GFP can be used to detect GFP by immunofluorescence microscopy in prokaryotic (E.coli) and eukaryotic (CHO cells) expression systems and detects GFP containing inserts. Significant amplification of signal is achieved using fluorochrome conjugated polyclonal anti-GFP relative to the fluorescence of GFP alone. For immunoblotting use either alkaline phosphatase or peroxidase conjugated polyclonal anti-GFP to detect GFP or GFP-containing proteins on western blots. Researchers should determine optimal titers for applications.

Purity/Specificity

GFP antibody was prepared from monospecific antiserum by immunoaffinity chromatography using Green Fluorescent Protein (Aequorea victoria) coupled to agarose beads followed by solid phase adsorption(s) to remove any unwanted reactivities.   Assay by immunoelectrophoresis resulted in a single precipitin arc against anti-Goat Serum and purified and partially purified Green Fluorescent Protein (Aequorea victoria).  No reaction was observed against Human, Mouse or Rat serum proteins.