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    hrp-gfp-antibody-ab6663.pdf

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Tags & Cell Markers Fusion / Marker Proteins GFP
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HRP Anti-GFP antibody (ab6663)

  • Datasheet
  • SDS
Submit a review Q&A (8)References (34)

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Abpromise

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Find out more.

Western blot - HRP Anti-GFP antibody (ab6663)

    Key features and details

    • HRP Goat polyclonal to GFP
    • Suitable for: WB
    • Reacts with: Species independent
    • Conjugation: HRP
    • Isotype: IgG

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    Overview

    • Product name

      HRP Anti-GFP antibody
      See all GFP primary antibodies
    • Description

      HRP Goat polyclonal to GFP
    • Host species

      Goat
    • Conjugation

      HRP
    • Tested applications

      Suitable for: WBmore details
    • Species reactivity

      Reacts with: Species independent
    • Immunogen

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

      MSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLKFICTT GKLPVPWPTL VTTFSYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDGNYKTRA EVKFEGDTLV NRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKVNFKIRHN IEDGSVQLAD HYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLEFVTAAGIT HGMDELYK


      Database link: P42212
      Run BLAST with BLAST the sequence with ExPASy Run BLAST with BLAST the sequence with NCBI
    • 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.

      Reproducibility is key to advancing scientific discovery and accelerating scientists’ next breakthrough.

      Abcam is leading the way with our range of recombinant antibodies, knockout-validated antibodies and knockout cell lines, all of which support improved reproducibility.

      We are also planning to innovate the way in which we present recommended applications and species on our product datasheets, so that only applications & species that have been tested in our own labs, our suppliers or by selected trusted collaborators are covered by our Abpromise™ guarantee.

      In preparation for this, we have started to update the applications & species that this product is Abpromise guaranteed for.

      We are also updating the applications & species that this product has been “predicted to work with,” however this information is not covered by our Abpromise guarantee.

      Applications & species from publications and Abreviews that have not been tested in our own labs or in those of our suppliers are not covered by the Abpromise guarantee.

      Please check that this product meets your needs before purchasing. If you have any questions, special requirements or concerns, please send us an inquiry and/or contact our Support team ahead of purchase. Recommended alternatives for this product can be found below, as well as customer reviews and Q&As.

    Properties

    • Form

      Liquid
    • Storage instructions

      Shipped at 4°C. Store at +4°C.
    • Storage buffer

      Preservative: 0.01% Gentamicin sulphate
      Constituents: 0.42% Potassium phosphate, 0.87% Sodium chloride, 1% 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

      • Tags & Cell Markers
      • Fusion / Marker Proteins
      • GFP

    Associated products

    • Recombinant Protein

      • Recombinant E. coli GFP protein (His tag) (ab119740)
      • Recombinant A. victoria GFP protein (ab84191)
    • Related Products

      • TMB ELISA Substrate (Highest Sensitivity) (ab171522)
      • TMB ELISA Substrate (High Sensitivity) (ab171523)
      • TMB ELISA Substrate (Fast Kinetic Rate) (ab171524)
      • TMB ELISA Substrate (Slow Kinetic Rate) (ab171525)
      • TMB ELISA Substrate (Slower Kinetic Rate) (ab171526)
      • TMB ELISA Substrate (Slowest Kinetic Rate) (ab171527)
      • 450 nm Stop Solution for TMB Substrate (ab171529)
      • 650 nm Stop Solution for TMB Substrate (ab171531)
      • Immunoassay Blocking Buffer (ab171534)
      • Immunoassay Blocking (BSA Free) (ab171535)
      • GFP ELISA Kit (ab171581)

    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

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

    Protocols

    • Western blot protocols

    Click here to view the general protocols

    Datasheets and documents

    • Datasheet
    • SDS
  • References (34)

    Publishing research using ab6663? Please let us know so that we can cite the reference in this datasheet.

    ab6663 has been referenced in 34 publications.

    • Fashe M  et al. Ser100-Phosphorylated RORa Orchestrates CAR and HNF4a to Form Active Chromatin Complex in Response to Phenobarbital to Regulate Induction of CYP2B6. Mol Pharmacol 97:191-201 (2020). PubMed: 31924695
    • Yi M  et al. Nuclear receptor CAR-ERa signaling regulates the estrogen sulfotransferase gene in the liver. Sci Rep 10:5001 (2020). PubMed: 32193417
    • Couturier L  et al. Regulation of Notch output dynamics via specific E(spl)-HLH factors during bristle patterning in Drosophila. Nat Commun 10:3486 (2019). PubMed: 31375669
    • Bouressam ML  et al. S-nitrosoglutathione inhibits cerebrovascular angiotensin II-dependent and -independent AT1 receptor responses: A possible role of S-nitrosation. Br J Pharmacol 176:2049-2062 (2019). PubMed: 30822355
    • Rogers C  et al. Gasdermin pores permeabilize mitochondria to augment caspase-3 activation during apoptosis and inflammasome activation. Nat Commun 10:1689 (2019). PubMed: 30976076
    View all Publications for this product

    Customer reviews and Q&As

    Show All Reviews Q&A
    Submit a review Submit a question

    1-8 of 8 Abreviews or Q&A

    Question

    Please let me know the exact molecular weight of the antibody. Is the antibody only the Fab fragment? What is the expected affinity between the antibody and GFP?

    Read More

    Abcam community

    Verified customer

    Asked on May 06 2006

    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.

    Read More

    Abcam Scientific Support

    Answered on May 08 2006

    Question

    About how many WB can I do with 1 vial? (= what is the initial volume and which dilution should I use?) Same question for ab 6673

    Read More

    Abcam community

    Verified customer

    Asked on Mar 04 2005

    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.

    Read More

    Abcam Scientific Support

    Answered on Mar 06 2005

    Question

    My GFP blot comes out w/a ton of background. I am using anti-GFP-ab6663 at 1:2000 in PBS/0.1%tween-20. Any suggestions?

    Read More

    Abcam community

    Verified customer

    Asked on Oct 28 2004

    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).

    Read More

    Abcam Scientific Support

    Answered on Oct 29 2004

    Question

    What was the source of GFP plates used for the ELISA?

    Read More

    Abcam community

    Verified customer

    Asked on Sep 10 2004

    Answer

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

    Read More

    Abcam Scientific Support

    Answered on Sep 13 2004

    Question

    Customer is interested in using this antibody in a sandwich ELISA and would like to know what concentration to use.

    Read More

    Abcam community

    Verified customer

    Asked on Sep 02 2004

    Answer

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

    Read More

    Abcam Scientific Support

    Answered on Sep 09 2004

    Question

    I was planning on using ab6663 to detect GFP+ cells in a paraffin-embedded organ (5um cut) from a transplanted mouse. i'll using DAB as a chromogen. i've had success using other biotinylated primary antibodies followed by HRP-Avidin then DAB. I was wondering if anyone else has used this product for this application and what concentration of ab6663 could be used. Thank you.

    Read More

    Abcam community

    Verified customer

    Asked on Oct 13 2003

    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.

    Read More

    Asdf Edo

    Abcam Scientific Support

    Answered on Oct 13 2003

    Question

    I was wondering if AB6663 is conjugated like a secondary antibody. If so, how can I use it in a sandwich ELISA? With what other antibody?

    Read More

    Abcam community

    Verified customer

    Asked on Feb 25 2002

    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.

    Read More

    Abcam Scientific Support

    Answered on Feb 26 2002

    Question

    Which of the three options for Abcam anti-GFP antibodies, rabbit, goat, mouse, which has the highest affinity? Thanks, Ann Vernallis

    Read More

    Abcam community

    Verified customer

    Asked on Jun 26 2001

    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.

    Read More

    Abcam Scientific Support

    Answered on Jun 27 2001

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