GFP Antibody (OARA01900)

Catalog No: OARA01900

Size:25UL

Western Blot of Anti-GFP (CHICKEN) antibody. Lane 1: MW. Lane 2: GFP. Load: 0.05 µg. Primary antibody: Anti-GFP (CHICKEN) antibody was used at 1:1000 overnight at 4C. Secondary antibody: Anti-Chicken IgG (GOAT) peroxidase conjugated antibody secondary antibody was used at 1:40,000 in Blocking Buffer for Fluorescent Western Blotting. Block: 1% BSA-TTBS (diluted to 1X) 30 min at 20C. 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 ug per lane. Primary antibody: GFP antibody at 2 ug/ml for 2 hr at room temperature. Secondary antibody: IRDye 800 Conjugated Affinity Purified anti-Chicken IgG [H&L] [Goat] MX10 at 1:20,000 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.

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.

Product Info

• Predicted Species Reactivity: Green Fluorescent Protein
• Product Format: Liquid (sterile filtered)
• Clonality: Polyclonal
• Host: Chicken
• Conjugation: Unconjugated
• Application: Dot blot|Enzyme-linked immunosorbent assay|Immunofluorescence|Immunohistochemistry|Multiplex assay|Purification|Western blot
• Reconstitution and Storage: Store Anti-GFP Antibody at -20C or below prior to opening. This vial contains a relatively low volume of reagent (25 μL). To minimize loss of volume dilute 1:10 by adding 225 μL of the buffer stated above directly to the vial. Recap, mix thoroughly and briefly centrifuge to collect the volume at the bottom of the vial. Use this intermediate dilution when calculating final dilutions as recommended below. Store the vial at -20C or below after dilution. Avoid cycles of freezing and thawing. Expiration date is three (3) months from date of opening.
• 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.
• Purification: Anti-GFP Antibody IgY purification 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.
• Concentration: 1.17 mg/ml
• Specificity: 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.
• Application Info: ELISA: 1:2,000~12,000
  IHC: 1:200~1,000
  WB: 1:200~1,000
  Optimal dilutions should be determined by the end user.
• Application Data: Anti-GFP is designed to detect GFP and its variants. 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 Aviva's monoclonal anti-GFP (OARA01901) 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 (OORA00141). 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.

Target Info

• Gene Symbol: GFP
• Gene Full Name: Green fluorescent protein
• Alias Symbols: chicken anti-GFP antibody, chicken Anti-Green Fluorescent Protein Antibody, GFP antibody, Green Fluorescent Protein antibody, EGFP, enhanced Green Fluorescent Protein, Aequorea victoria, Jellyfish anibody
• Protein Name: Green fluorescent protein
• Description of Target: Chicken Anti-GFP is designed for immunofluorescence microscopy, fluorescence based plate assays (FLISA) and fluorescent western blotting. GFP Antibody is also suitable for multiplex analysis, including multicolor imaging, utilizing various commercial platforms. Chicken IgY lacks the classic "Fc" domain and does not bind to mammalian IgG Fc receptors. Thus resulting in lower backgrounds for western blotting, ELISA and Immunohistochemistry.
• Uniprot ID: P42212