GFP antibody (Peroxidase)

• Catalog Number: orb345312
• Category: Antibodies
• Description: GFP antibody (Peroxidase)
• Species/Host: Goat
• Clonality: Polyclonal
• Tested applications: ELISA, IHC, WB
• Reactivity: Other
• Isotype: IgG
• Immunogen: Recombinant Green Fluorescent Protein (GFP) fusion protein corresponding to the full length amino acid sequence (246aa) derived from the jellyfish Aequorea victoria.
• Concentration: 1.0 mg/mL
• Dilution range: ELISA: 1:10,000 - 1:50,000, IHC: 1:500 - 1:2,000, WB: 1:2,000 - 1:5,000
• Form/Appearance: Lyophilized
• Purity: Anti-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, anti-Peroxidase and purified and partially purified Green Fluorescent Protein (Aequorea victoria). No reaction was observed against Human, Mouse and Rat Serum Proteins.
• Conjugation: HRP
• UniProt ID: P42212
• Storage: Store GFP antibody at 4° C prior to restoration. For extended storage aliquot contents and freeze at -20° C or below. Avoid cycles of freezing and thawing. Centrifuge product if not completely clear after standing at room temperature. This product is stable for several weeks at 4° C as an undiluted liquid. Dilute only prior to immediate use.
• Buffer/Preservatives: 0.01% (w/v) Gentamicin Sulfate. Do NOT add Sodium Azide!
• Alternative names: goat anti-GFP Antibody peroxidase Conjugation, HRP
• Note: For research use only
• Application notes: Polyclonal anti-GFP is designed to detect GFP and its variants. Anti-GFP Peroxidase conjugated antibody has been tested by ELISA to detect GFP by ELISA (sandwich or capture) for the direct binding of antigen and recognizes wild type, recombinant and enhanced forms of GFP and by western blot. 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 conjugated 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 can detect 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. Optimal titers for applications should be determined by the researcher.
• Expiration Date: 12 months from date of receipt.

Western Blot of Goat anti-GFP antibody Peroxidase conjugated. Marker: Opal Pre-stained ladder. Lane 2: HEK293 lysate (p/n orb348669). Lane 3: HeLa Lysate (p/n orb348667). Lane 4: CHO/K1 Lysate. Lane 5: C2C12 Lysate. Lane 6: NIH/3T3 Lysate (p/n orb348714). Lane 7: Mouse Embryonic Fibroblast Lysate (p/n orb348722). Lane 8: E-coli HCP Control (p/n orb342262). Lane 9: 2 Epitope MBP Tag Marker Lysate (p/n orb348609). Lane 10: Recombinant Red Fluorescent Protein (p/n orb345960). Lane 11: Green Fluorescent Protein (p/n orb345957). Lane 12: Glutathione-S-Transferase Protein. Lane 13: Maltose Binding Protein. Marker: Opal Pre-stained ladder Load: 10 µg of lysate or 50 ng of purified protein per lane. Primary antibody: goat anti-GFP peroxidase conjugated antibody at 1:1000 overnight at 4C. Secondary antibody: none Blocking Buffer: BlockOut (p/n orb348644) for 30 min at RT. Predicted/Observed size: 28 kDa for GFP.