Green Fluorescent Protein (GFP) (AP)

Designed to detect GFP and its variants. Monoclonal and polyclonal forms of anti-GFP were assayed by ELISA (sandwich or capture) for direct binding of antigen and recognize wild type, recombinant and enhanced forms of GFP. Biotin conjugated monoclonal and polyclonal forms anti-GFP were assayed in a sandwich ELISA are well suited to titrate GFP in solution using either form of the antibody as the capture or detection antibodies. The detection antibody is typically conjugated to biotin and subsequently reacted with streptavidin-HRP. Fluorochrome conjugated polyclonal anti-GFP was assayed by immuno-fluorescence microscopy on prokaryotic (E.coli) and eukaryotic (CHO cells) expression systems and was shown to detect GFP containing inserts. Significant amplification of signal was detected using fluorochrome conjugated polyclonal anti-GFP relative to the fluorescence of GFP alone. Alkaline Phosphatase or Peroxidase conjugated polyclonal anti-GFP assayed by immunoblot shows a 42kD band when reacted with GFP on a western blot.

Applications:
Suitable for use in ELISA, Western Blot and Immunohistochemistry. Other applications not tested.

Recommended Dilutions:
ELISA: 1:16,000-1:32,000, 1.0 μg of GFP in a standard capture ELISA using pNPP p-nitrophenyl phosphate code as a substrate for 30 minutes at room temperature.
Optimal dilution determined by the researcher.

Label: Alkaline Phosphatase (Calf Intestine) (Molecular Weight 140,000 daltons)

Storage and Stability:
May be stored at 4°C for short-term only. For long-term storage, store at -20°C. Aliquots are stable for at least 6 months at -20°C. For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap. Further dilutions can be made in assay buffer.