GFP Antibody

SKU: orb345912

Description

GFP antibody

Images & Validation

• Tested Applications: ELISA, IHC, WB
• Dilution range: ELISA: 1:20,000 - 1:120,000, IHC: 1:200 - 1:1,000, WB: 1:1,000 - 1:5,000
• Reactivity: Other
• Application Notes:
  Polyclonal anti-GFP is designed to detect GFP and its variants. Anti-GFP antibody has been tested by ELISA and western blot and is suitable for immunofluorescence. 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 Biorbyt's monoclonal anti-GFP using either form of the antibody as the capture or detection antibodies. 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. 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.

Key Properties

• Antibody Type: Primary Antibody
• Host: Gallus
• Clonality: Polyclonal
• Isotype: IgY
• Immunogen: The immunogen is a Green Fluorescent Protein (GFP) fusion protein corresponding to the full length amino acid sequence (246aa) derived from the jellyfish Aequorea victoria.
• Purity: Anti-GFP Antibody was prepared from egg yolks 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-Chicken Serum and purified and partially purified Green Fluorescent Protein (Aequorea victoria). No reaction was observed against Human, Mouse or Rat serum proteins.

Storage & Handling

• Storage: Store GFP Antibody at -20° C prior to opening. Aliquot contents and freeze at -20° C or below for extended storage. 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.
• Form/Appearance: Liquid (sterile filtered)
• Buffer/Preservatives: 0.01% (w/v) Sodium Azide
• Concentration: 1.16 mg/mL
• Dry Ice Shipping: Please note: This product requires shipment on dry ice. A dry ice surcharge will apply.
• Disclaimer: For research use only

Alternative Names

chicken anti-GFP antibody, Green Fluorescent Protein, GFP antibody, Green Fluorescent Protein antibody, EGFP, enhanced Green Fluorescent Protein, Aequorea victoria, Jellyfish

Immunofluorescence Microscopy of Chicken anti-GFP antibody. Tissue: KruppleGAL4 driver line in Drosophila eye disc. Fixation: 0.5% PFA. Antigen retrieval: not required. Primary antibody: anti-GFP antibody diluted 1:500 for 2 hr at RT. Secondary antibody: Alexa™488 conjugated anti-Chicken IgG at 1:300 for 1 hr at RT. Blocking: 5% NGS in PBS with 0.1% Triton X-100 for 15 min. Localization: E2F-1 pS364 is nuclear and occasionally cytoplasmic. Staining: recombinant tau-myc-GFP protein as green fluorescent signal.

Western Blot of Anti-GFP (CHICKEN) antibody. Lane 1: MW. Lane 2: GFP (p/n orb345957) Load: 0.05 µg. Primary antibody: Anti-GFP (CHICKEN) antibody (p/n orb345912) was used at 1:1000 overnight at 4°C. Secondary antibody: Anti-Chicken IgG (GOAT) peroxidase conjugated (p/n orb346892) secondary antibody was used at 1:40000 in Blocking Buffer for Fluorescent Western Blotting (p/n orb348637). Block: 1% BSA-TTBS (p/n orb348540, diluted to 1X) 30 min at 20°C. Predicted/Observed size: 27 kDa for GFP. Other band(s): none.

Western Blot of Anti-GFP antibody. Lane 1: mouse spleen lysate. Lane 2: mouse spleen lysate spiked with 50 ng of wt GFP. Load: 20 µg per lane. Primary antibody: GFP antibody at 2 µg/ml for 2 hr at room temperature. Secondary antibody: IRDye™800 Conjugated Affinity Purified anti-Chicken IgG [H&L] [Goat] MX10 at 1:20000 for 45 min at RT. Block: 5% BSA in PBS 2 hr at room temperature. Predicted/Observed size: 27 kDa for GFP epitope tag. Other band(s): none.

Western Blot Results of Chicken Anti-GFP Antibody. Lane 1: Opal Prestained Molecular Weight Marker. Lane 2: GFP (p/n orb345957) / HeLa WCL (p/n orb348668) [0.05 µg/10 µg]. Lane 3: GFP (p/n orb345957) / NIH3T3 WCL [0.05 µg/10 µg]. Lane 4: GFP (p/n orb345957) / PC-12 WCL [0.05 µg/10 µg]. Lane 5: HeLa WCL (p/n orb348668) [10 µg]. Lane 6: NIH3T3 WCL [10 µg]. Lane 7: PC-12 WCL [10 µg]. Primary Antibody: Anti-GFP at 1.0 µg/ml overnight at 2-8°C. Secondary Antibody: Goat Anti-Chicken IgG HRP (p/n orb346892) at 1:40000 for 30 mins at RT. Block: orb348637 Buffer for 1hr at RT. Predicted MW: 27kda. Exposure: 2 seconds.