Recombinant
RabMAb

Anti-GFP antibody [EPR14104] - BSA and Azide free (ab220802)

Overview

  • Product name
    Anti-GFP antibody [EPR14104] - BSA and Azide free
    See all GFP primary antibodies
  • Description
    Rabbit monoclonal [EPR14104] to GFP - BSA and Azide free
  • Host species
    Rabbit
  • Tested applications
    Suitable for: Flow Cyt, ICC/IF, WB, IHC-Pmore 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

    The formulation and the concentration of this product is compatible for metal-conjugation for mass cytometry (CyTOF®). 

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

    Use our conjugation kits for antibody conjugates that are ready-to-use in as little as 20 minutes with <1 minute hands-on-time and 100% antibody recovery: available for fluorescent dyes, HRP, biotin and gold.

    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 = 1.11 x 10 -11 M
    Learn more about KD
  • Storage buffer
    Constituent: PBS
  • Concentration information loading...
  • Purity
    Protein A purified
  • Clonality
    Monoclonal
  • Clone number
    EPR14104
  • Isotype
    IgG
  • Research areas

Applications

Our Abpromise guarantee covers the use of ab220802 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
Flow Cyt Use at an assay dependent concentration.
ICC/IF Use at an assay dependent concentration.
WB Use at an assay dependent concentration. Detects a band of approximately 27 kDa (predicted molecular weight: 27 kDa).
IHC-P Use at an assay dependent concentration. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

See IHC antigen retrieval protocols.

  • 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

    • 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).

      This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab183734).

    • 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.

      This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab183734).

    • 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.

      This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab183734).

    • 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.

      This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab183734).

    • 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.

      This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab183734).

    • 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.

      This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab183734).

    • 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)

      This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab183734).

    References

    ab220802 has not yet been referenced specifically in any publications.

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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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