Overview

  • Product name
    Anti-GFP antibody [EPR14104]
    See all GFP primary antibodies
  • Description
    Rabbit monoclonal [EPR14104] to GFP
  • Host species
    Rabbit
  • Tested applications
    Suitable for: WB, IHC-P, ICC/IF, Flow Cytmore details
    Unsuitable for: IP
  • 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
    • WB: GFP transfected 293 cell lysate. IHC-P: GFP transgenic mouse colon tissue and GFP transgenic mouse liver tissue. ICC/IF: GFP transfected 293 cells. ICC-IF: GFP transfected NIH3T3 cells.
  • General notes

    On the basis of low sequence homology, ab183734 is predicted to show no or limited cross-reactivity to RFP, BFP, and CFP.

    For the best results in IHC, please use ab183734 on Formalin/PFA-fixed paraffin-embedded sections with heat mediated antigen retrieval.

    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.

    We are constantly working hard to ensure we provide our customers with best in class antibodies. As a result of this work we are pleased to now offer this antibody in purified format. We are in the process of updating our datasheets. The purified format is designated 'PUR' on our product labels. If you have any questions regarding this update, please contact our Scientific Support team.

    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 = 1.11 x 10 -11 M
    Learn more about KD
  • Storage buffer
    Preservative: 0.01% Sodium azide
    Constituents: 59% PBS, 40% Glycerol, 0.05% BSA
  • Concentration information loading...
  • Purity
    Protein A purified
  • Clonality
    Monoclonal
  • Clone number
    EPR14104
  • Isotype
    IgG
  • Research areas

Applications

Our Abpromise guarantee covers the use of ab183734 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 1/10000 - 1/50000. Detects a band of approximately 27 kDa (predicted molecular weight: 27 kDa).
IHC-P 1/100 - 1/250. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

See IHC antigen retrieval protocols.

ICC/IF 1/500.
Flow Cyt Use at an assay dependent concentration.
  • Application notes
    Is unsuitable for IP.
  • 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

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of GFP transgenic mouse pancreas tissue labelling GFP with purified ab183734 at a dilution of 1/100. Heat mediated antigen retrieval was performed using Tris/EDTA buffer pH 9. ab97051, a HRP-conjugated goat anti-rabbit IgG (H+L) was used as the secondary antibody (1/500). Negative control using PBS instead of primary antibody. Counterstained with hematoxylin.

    • ab183734 staining GFP in GFP-transfected NIH3T3 cells. The cells were fixed with 4% formaldehyde (10min) and then blocked in 1% BSA / 0.3M glycine in 0.1%PBS-Tween for 1h. The cells were then incubated with ab183734 at 1/500 dilution overnight at +4°C followed by incubation with ab150081, Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488), for 1 hour, at 1μg/ml.

      Under identical experimental conditions, when compared to the basal level of GFP expression in transfected NIH3T3 cells, the cells upon which ab183734 was applied gave a stronger signal in the 488 channel, indicating that ab183734 is binding to GFP and therefore eliciting signal amplification.

      ab183734 was also applied to non-GFP-transfected NIH3T3 cells, which produced no positive staining, indicating specificity for GFP. Nuclear DNA was labelled with 1.43μM DAPI (blue).

    • All lanes : Anti-GFP antibody [EPR14104] (ab183734) at 1/10000 dilution (purified)

      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 (HRP) (ab97051) at 1/100000 dilution

      Predicted band size: 27 kDa
      Observed band size: 25 kDa
      why is the actual band size different from the predicted?



      Blocking and dilution buffer: 5% NFDM /TBST.

    • Flow Cytometry analysis of 293T (human embryonic kidney) transfected with human TNFRSF9 cells labeling GFP with purified ab183734 at 1/50 dilution(10ug/ml) (red). 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 (Black) was used as the isotype control, cells without incubation with primary antibody and secondary antibody (Blue) was used as the unlabeled control.

    • Immunohistochemical analysis of paraffin-embedded GFP transgenic mouse colon tissue (left) and normal mouse colon tissue (right) labeling GFP with unpurified ab183734 at 1/250 dilution followed by prediluted HRP Polymer for Rabbit IgG. Counterstained with Hematoxylin.

    • Immunofluorescent analysis of 4% paraformaldehyde-fixed GFP transfected 293 cells labeling GFP with unpurified ab183734 at 1/500 dilution, followed by Goat anti rabbit IgG (Alexa Fluor® 488) secondary antibody at 1/200 dilution (green). Counterstained with Dapi (blue)

    • All lanes : Anti-GFP antibody [EPR14104] (ab183734) at 1/10000 dilution (unpurified)

      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

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of mouse pancreas tissue (negative control) labelling GFP with purified ab183734 at a dilution of 1/100. Heat mediated antigen retrieval was performed using Tris/EDTA buffer pH 9. ab97051, a HRP-conjugated goat anti-rabbit IgG (H+L) was used as the secondary antibody (1/500). Negative control using PBS instead of primary antibody. Counterstained with hematoxylin.

    • Immunohistochemical analysis of paraffin-embedded GFP transgenic mouse liver tissue (left) and normal mouse liver tissue (right) labeling GFP with unpurified ab183734 at 1/250 dilution followed by prediluted HRP Polymer for Rabbit IgG. Counterstained with Hematoxylin.

    References

    This product has been referenced in:
    • Guadagnin E  et al. Nuclear localized Akt limits skeletal muscle derived fibrotic signaling. Biochem Biophys Res Commun 508:838-843 (2019). Read more (PubMed: 30528731) »
    • Counsell JR  et al. Foamy Virus Vectors Transduce Visceral Organs and Hippocampal Structures following In Vivo Delivery to Neonatal Mice. Mol Ther Nucleic Acids 12:626-634 (2018). Read more (PubMed: 30081233) »
    See all 8 Publications for this product

    Customer reviews and Q&As

    1-10 of 22 Abreviews or Q&A

    Abcam guarantees this product to work in the species/application used in this Abreview.
    Application
    Immunocytochemistry/ Immunofluorescence
    Sample
    Mouse Cell (MC3T3)
    Permeabilization
    Yes - NP40
    Specification
    MC3T3
    Blocking step
    BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 3% · Temperature: 21°C
    Fixative
    Formaldehyde

    Abcam user community

    Verified customer

    Submitted Feb 19 2019

    Abcam guarantees this product to work in the species/application used in this Abreview.
    Application
    Western blot
    Sample
    Mouse Cell lysate - whole cell (MC3T3)
    Gel Running Conditions
    Reduced Denaturing
    Loading amount
    20 µg
    Treatment
    Transfected with GFP
    Specification
    MC3T3
    Blocking step
    Milk as blocking agent for 2 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 21°C

    Abcam user community

    Verified customer

    Submitted Feb 19 2019

    Abcam guarantees this product to work in the species/application used in this Abreview.
    Application
    Western blot
    Sample
    Human Cell lysate - whole cell (U2OS)
    Gel Running Conditions
    Reduced Denaturing
    Loading amount
    15 µg
    Specification
    U2OS
    Blocking step
    Milk as blocking agent for 2 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 21°C

    Abcam user community

    Verified customer

    Submitted Jan 28 2019

    Abcam guarantees this product to work in the species/application used in this Abreview.
    Application
    Immunocytochemistry/ Immunofluorescence
    Sample
    Human Cell (U2OS)
    Permeabilization
    Yes - NP40
    Specification
    U2OS
    Blocking step
    BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 3% · Temperature: 21°C
    Fixative
    Formaldehyde

    Abcam user community

    Verified customer

    Submitted Jan 28 2019

    Abcam guarantees this product to work in the species/application used in this Abreview.
    Application
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
    Sample
    Mouse Tissue sections (Muscle)
    Antigen retrieval step
    Heat mediated - Buffer/Enzyme Used: pH 9.0 EDTA
    Permeabilization
    No
    Specification
    Muscle
    Blocking step
    GeneTex GTX48880 as blocking agent for 10 minute(s) · Concentration: 1% · Temperature: RT°C
    Fixative
    Formaldehyde

    Abcam user community

    Verified customer

    Submitted May 10 2018

    Application
    Immunocytochemistry
    Sample
    Dog Cultured Cells (GFP-transduced neural precursor cells)
    Permeabilization
    Yes - 0.5% TritonX
    Specification
    GFP-transduced neural precursor cells
    Blocking step
    BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 10% · Temperature: 25°C
    Fixative
    Paraformaldehyde

    Abcam user community

    Verified customer

    Submitted May 09 2018

    Abcam guarantees this product to work in the species/application used in this Abreview.
    Application
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
    Sample
    Rat Tissue sections (Liver)
    Antigen retrieval step
    Heat mediated - Buffer/Enzyme Used: pH 9.0 EDTA
    Permeabilization
    No
    Specification
    Liver
    Fixative
    Formaldehyde

    Abcam user community

    Verified customer

    Submitted Jan 04 2018

    Application
    Immunohistochemistry free floating
    Sample
    Mouse Tissue sections (Spinal Cord Dorsal Horn)
    Specification
    Spinal Cord Dorsal Horn

    Steven West

    Verified customer

    Submitted Oct 27 2017

    Application
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
    Sample
    Mouse Tissue sections (mouse tissue section (skin))
    Antigen retrieval step
    Heat mediated - Buffer/Enzyme Used: Citrate Buffer pH 6.0
    Permeabilization
    Yes - 0.3% triton in PBS
    Specification
    mouse tissue section (skin)
    Blocking step
    Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
    Fixative
    Paraformaldehyde

    Dr. Yulia Shwartz

    Verified customer

    Submitted Aug 17 2017

    Abcam guarantees this product to work in the species/application used in this Abreview.
    Application
    Western blot
    Sample
    Mouse Tissue lysate - whole (various mouse tissue lysate: skeletal muscle, hear)
    Gel Running Conditions
    Non-reduced Denaturing (10% gel)
    Loading amount
    40 µg
    Specification
    various mouse tissue lysate: skeletal muscle, hear
    Blocking step
    Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 18°C

    Abcam user community

    Verified customer

    Submitted Jun 19 2017

    1-10 of 22 Abreviews or Q&A

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