Overview

  • Product name
    Anti-GFP antibody (HRP)
    See all GFP primary antibodies
  • Description
    Goat polyclonal to GFP (HRP)
  • Host species
    Goat
  • Conjugation
    HRP
  • Tested applications
    Suitable for: WBmore details
  • Immunogen

    Recombinant full length protein corresponding to GFP aa 1-246.
    Sequence:

    MSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLKFICTT GKLPVPWPTL VTTFSYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDGNYKTRA EVKFEGDTLV NRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKVNFKIRHN IEDGSVQLAD HYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLEFVTAAGIT HGMDELYK


    Database link: P42212

  • Positive control
    • GFP
  • General notes

    Designed to detect GFP and its variants in ELISA (sandwich or capture), immunoblotting and immunoprecipitation Peroxidase conjugated anti-GFP assayed by immunoblot shows a 42 kDa band when reacted with GFP on a western blot.

Properties

  • Form
    Liquid
  • Storage instructions
    Shipped at 4°C. Store at +4°C.
  • Storage buffer
    pH: 7.20
    Preservative: 0.01% Gentamicin sulphate
    Constituents: 0.42% Potassium phosphate, 0.87% Sodium chloride, 99% BSA

    Do NOT add Sodium Azide!
  • Concentration information loading...
  • Purity
    Affinity purified
  • Purification notes
    This product 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.
  • Primary antibody notes
    Designed to detect GFP and its variants in ELISA (sandwich or capture), immunoblotting and immunoprecipitation Peroxidase conjugated anti-GFP assayed by immunoblot shows a 42 kDa band when reacted with GFP on a western blot.
  • Clonality
    Polyclonal
  • Isotype
    IgG
  • Research areas

Applications

Our Abpromise guarantee covers the use of ab6663 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Application Abreviews Notes
WB 1/2000 - 1/5000.

Target

  • Relevance
    Function: Energy-transfer acceptor. Its role is to transduce the blue chemiluminescence of the protein aequorin into green fluorescent light by energy transfer. Fluoresces in vivo upon receiving energy from the Ca2+ -activated photoprotein aequorin.

    Subunit structure: Monomer.

    Tissue specificity: Photocytes.

    Post-translational modification: Contains a chromophore consisting of modified amino acid residues. The chromophore is formed by autocatalytic backbone condensation between Ser-65 and Gly-67, and oxidation of Tyr-66 to didehydrotyrosine. Maturation of the chromophore requires nothing other than molecular oxygen.

    Biotechnological use: Green fluorescent protein has been engineered to produce a vast number of variously colored mutants, fusion proteins, and biosensors. Fluorescent proteins and its mutated allelic forms, blue, cyan and yellow have become a useful and ubiquitous tool for making chimeric proteins, where they function as a fluorescent protein tag. Typically they tolerate N- and C-terminal fusion to a broad variety of proteins. They have been expressed in most known cell types and are used as a noninvasive fluorescent marker in living cells and organisms. They enable a wide range of applications where they have functioned as a cell lineage tracer, reporter of gene expression, or as a measure of protein-protein interactions. Can also be used as a molecular thermometer, allowing accurate temperature measurements in fluids. The measurement process relies on the detection of the blinking of GFP using fluorescence correlation spectroscopy.

    Sequence similarities: Belongs to the GFP family.

    Biophysicochemical properties: Absorption: Abs(max)=395 nm
    Exhibits a smaller absorbance peak at 470 nm. The fluorescence emission spectrum peaks at 509 nm with a shoulder at 540 nm.
  • Alternative names
    • GFP antibody
    • Green fluorescent protein antibody

Images

  • Anti-GFP antibody (HRP) (ab6663) + GFP

References

This product has been referenced in:
  • Shizu R  et al. Phosphorylated Nuclear Receptor CAR Forms a Homodimer To Repress Its Constitutive Activity for Ligand Activation. Mol Cell Biol 37:N/A (2017). Read more (PubMed: 28265001) »
  • Couturier L  et al. Regulation of cortical stability by RhoGEF3 in mitotic Sensory Organ Precursor cells inDrosophila. Biol Open 6:1851-1860 (2017). Read more (PubMed: 29101098) »
See all 23 Publications for this product

Customer reviews and Q&As

1-8 of 8 Abreviews or Q&A

Answer

Thank you for your enquiry. To answer your questions: Purified GFP is a 27 kDa monomer consisting of 238 amino acids. Ab6663 GFP antibody is whole IgG molecule. Unfortunately we do not have details of the affinities between ab6663 and its antigen.

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Answer

The number of Western Blots will depend on the volume of diluent and dilution used. I would suggest trying a preliminary experiment doing a dilution curve with a range of concentrations (1:500-1:5000 for example) and optimising from there. If you use 10 ml of buffer and 1:500 dilution you will get 5 blots from the vial, at 1:5000 you will get 50.

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Answer

At this point I would suggest the following to help decrease the amount of background you are seeing. Try blocking with 3% BSA. Incubate with ab6663 for 1 hour at RT rather than overnight at 4C. Also, increase the number of your washes and you may want to decrease the concentration of the primary even more (try 1:2500).

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Question
Answer

Our Elisa plates are usually from Thermo Electron. We have our in-house recipe of diluent.

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Answer

Although this antibody is not tested in ELISA, I would recommend using a concentration of 1-10 ug/mL.

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Answer

To our knowledge, this antibody has yet to be tested in this application. All tested applications are specified on Abcam product datasheets. If you decide to go ahead and purchase this product, please let us know how you get on and in return we will forward a reward of your choice, typically an Amazon gift voucher.

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Answer

ab6663 is not conjugated to permit direct detection in a sandwich ELISA. You must use an anti goat conjugated secondary to detect this polyclonal goat primary antibody.

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Answer

We do not have specific affinity constant values for these antibodies but the one that is used most routinely and is most recommended is ab290.

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Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"

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