Overview

  • Product name
    Anti-GFP antibody [EPR14104-89]
    See all GFP primary antibodies
  • Description
    Rabbit monoclonal [EPR14104-89] to GFP
  • Host species
    Rabbit
  • Tested applications
    Suitable for: ICC/IF, IHC-P, WB, Flow Cytmore details
  • Species reactivity
    Reacts with: Species independent
  • Immunogen

    Full length protein corresponding to Aequorea victoria GFP aa 1 to the C-terminus.
    Database link: P42212

  • Positive control
    • GFP transfected 293 cell lysate; GFP transgenic mouse colon tissue; GFP transgenic mouse liver tissue; GFP transfected 293 cells.
  • General notes

    On the basis of low sequence homology, ab183735 is predicted to show no or limited cross-reactivity to RFP, YFP, BFP, and CFP.

    A trial size is available to purchase for this antibody.

    Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMab® patents.

    This product is a recombinant rabbit monoclonal antibody.

Properties

  • Form
    Liquid
  • Storage instructions
    Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C long term. Avoid freeze / thaw cycle.
  • Dissociation constant (KD)
    KD = 8.82 x 10 -11 M
    Learn more about KD
  • Storage buffer
    Preservative: 0.01% Sodium azide
    Constituents: 59% PBS, 40% Glycerol, 0.05% BSA
  • Purity
    Tissue culture supernatant
  • Clonality
    Monoclonal
  • Clone number
    EPR14104-89
  • Isotype
    IgG
  • Research areas

Applications

Our Abpromise guarantee covers the use of ab183735 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
ICC/IF 1/500.
IHC-P 1/250. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
WB 1/10000 - 1/50000. Detects a band of approximately 27 kDa (predicted molecular weight: 27 kDa).
Flow Cyt Use at an assay dependent concentration.

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

  • Immunohistochemical analysis of paraffin-embedded GFP transgenic mouse colon tissue (left) and normal mouse colon tissue (right) labeling GFP with ab183735 at 1/250 dilution followed by prediluted HRP Polymer for Rabbit IgG. Counter stained with Hematoxylin.

  • All lanes : Anti-GFP antibody [EPR14104-89] (ab183735) at 1/10000 dilution

    Lane 1 : GFP transfected 293 cell lysate
    Lane 2 : Non-transfected 293 cell lysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugate at 1/1000 dilution

    Predicted band size: 27 kDa
    Observed band size: 27 kDa

  • Immunofluorescent analysis of 4% paraformaldehyde-fixed GFP transfected 293 cells labeling GFP with ab183735 at 1/500 dilution, followed by Goat anti rabbit IgG (Alexa Fluor® 488) secondary antibody at 1/200 dilution (green). Counter stained with Dapi (blue).

  • Immunohistochemical analysis of paraffin-embedded GFP transgenic mouse liver tissue (left) and normal mouse liver tissue (right) labeling GFP with ab183735 at 1/250 dilution followed by prediluted HRP Polymer for Rabbit IgG. Counter stained with Hematoxylin.

  • Flow Cytometry analysis of 293T (Human epithelial cell line from embryonic kidney) transfected with GFP cells labeling GFP with unpurified ab183735 at 1/200 dilution (10ug/ml, Right panel). Cells were fixed with 4% paraformaldehyde and permeabilised with 90% methanol. A Goat anti rabbit IgG (Alexa Fluor® 647)(1/2000 dilution) was used as the secondary antibody. Rabbit monoclonal IgG (Left panel) was used as the isotype control.

References

This product has been referenced in:
  • Zhu M  et al. Transplantation of periodontal ligament cell sheets expressing human ß-defensin-3 promotes anti-inflammation in a canine model of periodontitis. Mol Med Rep 16:7459-7467 (2017). Read more (PubMed: 28944821) »
  • Mukai A  et al. Reliable handling of highly A/T-rich genomic DNA for efficient generation of knockin strains of Dictyostelium discoideum. BMC Biotechnol 16:37 (2016). Read more (PubMed: 27075750) »
See all 2 Publications for this product

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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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