Overview

  • Product name
  • Description
    Goat polyclonal to GFP
  • Host species
    Goat
  • Specificity
    Anti-GFP assayed by ELISA for direct binding of antigen recognizes wild type, recombinant and enhanced forms of GFP.
  • Tested applications
    Suitable for: WB, IP, ELISA, ICC/IF, IHC-P, IHC-FrFl, IHC-Frmore details
  • Immunogen

    Fusion protein corresponding to Aequorea victoria GFP aa 1-246.
    Database link: P42212

  • Positive control
    • HeLa nuclear extract lysate (ab150036) can be used as a positive control in WB. E5.5 Hex-GFP transgenic mouse embryo.

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. Avoid freeze / thaw cycle.
  • Storage buffer
    pH: 7.20
    Preservative: 0.01% Sodium azide
    Constituents: 0.42% Potassium phosphate, 0.87% Sodium chloride
  • 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.
  • Clonality
    Polyclonal
  • Isotype
    IgG
  • Research areas

Applications

Our Abpromise guarantee covers the use of ab6673 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 1/400 - 1/2000.

(for immunoprecipitated GFP, see Abreview).

IP Use at an assay dependent concentration.
ELISA 1/40000.

This antibody can be used to detect GFP by ELISA (sandwich or capture) for the direct binding of antigen and recognizes wild type, recombinant and enhanced forms of GFP.

ICC/IF 1/500.
IHC-P 1/200 - 1/1000.
IHC-FrFl Use at an assay dependent concentration.
IHC-Fr 1/200 - 1/1000.

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

  • Immunofluorescence of TGN mouse liver labeling GFP on hepatocytes with ab6673.

  • All lanes : Anti-GFP antibody (ab6673) at 1 µg/ml

    Lane 1 : HeLa cells
    Lane 2 : mock transfected HeLa cell lysate

    Lysates/proteins at 35 µg per lane.

    Additional bands at: 33 kDa. We are unsure as to the identity of these extra bands.

  • All lanes : Anti-GFP antibody (ab6673) at 1/1000 dilution

    Lane 1 : MRC5VA lung fibroblast whole cell lysate overexpressing EGFP alone
    Lanes 2-3 : MRC5VA lung fibroblast whole cell lysate overexpressing an EGFP fusion protein

    Lysates/proteins at 15 µg per lane.

    Secondary
    All lanes : HRP-conjugated anti-goat polyclonal at 1/10000 dilution

    Developed using the ECL technique.

    Performed under reducing conditions.

    Observed band size: 27,55 kDa
    why is the actual band size different from the predicted?


    Exposure time: 5 seconds

    See Abreview

  • Immunohistochemistry of GFP transgenic mouse liver labeling GFP with ab6673. 

References

This product has been referenced in:
  • Goldman SM  et al. Co-delivery of a laminin-111 supplemented hyaluronic acid based hydrogel with minced muscle graft in the treatment of volumetric muscle loss injury. PLoS One 13:e0191245 (2018). Read more (PubMed: 29329332) »
  • Campbell K  et al. Differential roles of the Drosophila EMT-inducing transcription factors Snail and Serpent in driving primary tumour growth. PLoS Genet 14:e1007167 (2018). Read more (PubMed: 29420531) »
See all 203 Publications for this product

Customer reviews and Q&As

1-10 of 22 Abreviews or Q&A

Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Blocking step
BSA as blocking agent for 40 minute(s) · Concentration: 1% · Temperature: RT°C
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: citrate buffer pH6
Sample
Mouse Tissue sections (spleen, red pulp, hematopoietic tissue)
Specification
spleen, red pulp, hematopoietic tissue
Permeabilization
Yes - Tween-20
Fixative
Paraformaldehyde

Abcam user community

Verified customer

Submitted Aug 01 2014

Application
Immunohistochemistry (PFA perfusion fixed frozen sections)
Blocking step
Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: RT°C
Antigen retrieval step
None
Sample
Mouse Tissue sections (Brain, Neural Stem Cells)
Specification
Brain, Neural Stem Cells
Permeabilization
No
Fixative
Paraformaldehyde

Miss. Katya Zelentsova

Verified customer

Submitted Feb 26 2014

Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Mouse Tissue sections (GFP labelled 4T1 cells in the lung)
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: ph9
Permeabilization
No
Specification
GFP labelled 4T1 cells in the lung
Blocking step
BSA as blocking agent for 30 minute(s) · Concentration: 1% · Temperature: 25°C
Fixative
Formaldehyde

Dr. Victoria Bridgeman

Verified customer

Submitted May 27 2016

Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Mouse Tissue sections (esophagus)
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: 10mM citric acid, pH6.0
Specification
esophagus
Blocking step
Non-Abcam blocking agent as blocking agent for 20 minute(s) · Concentration: 100% · Temperature: 20°C
Fixative
Formaldehyde

Tatiana Karakasheva

Verified customer

Submitted Feb 22 2016

Question
Answer

This anti GFP antibody has reacted with all tested variants of GFP from A. Victoria tested including YFP. We have not tested with eYFP specifically. The lots we have tested have not shown cross reactivity to RFP.

Read More
Application
IHC - Wholemount
Sample
Mouse Tissue (Epidermal sheet of the skin)
Specification
Epidermal sheet of the skin

Miss. yaara tabib

Verified customer

Submitted Jun 19 2014

Application
Immunohistochemistry (Frozen sections)
Blocking step
BSA as blocking agent for 20 minute(s) · Concentration: 1% · Temperature: 22°C
Sample
Mouse Tissue sections (whole mouse embryo)
Specification
whole mouse embryo
Permeabilization
Yes - 0.1% Triton-X 100 in TBS
Fixative
Paraformaldehyde

Abcam user community

Verified customer

Submitted May 31 2014

Abreviews
Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Western blot
Loading amount
20 µg
Gel Running Conditions
Non-reduced Denaturing (10)
Sample
Human Cell lysate - whole cell (HeLa cells)
Specification
HeLa cells
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 21°C

Abcam user community

Verified customer

Submitted May 28 2013

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 (Brain)
Specification
Brain
Fixative
Paraformaldehyde
Antigen retrieval step
None
Permeabilization
Yes - Triton X-100 (0.3%) in blocking buffer
Blocking step
Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 10% · Temperature: RT°C

Abcam user community

Verified customer

Submitted Dec 19 2012

Answer

Gracias por contactarnos.

El anticuerpo ab6673 se ha testada en inmunofluorescencia con tejidos, bien sea congelados, o embebidos en parafina. No tenemos constancia de su uso en células, por lo que si esta es la aplicación que queréis llevar a cabo, te animaría a que me lo comentaras, y quizás podamos encontrar otra alternativa que ya haya sido testada, y que por tanto esté garantizada.

En general recomendamos conservar los anticuerpos a 4C si se consumen en la semana o dos semanas siguientes a su recibimiento, aunque en principio a 4C son estables hasta un año. A -20C se estima que se mantengan estables hasta 5 años. En cualquier caso se aconseja alicuotarlo en cuanto llegue y evitar ciclos de congelación / descongelación.

De cualquier manera, os animamos a usar el anticuerpo en los 6 meses siguientes a la fecha de compra, para en caso de tener cualquier problema con él, que éste siga estando bajo nuestra garantía https://www.abcam.com/index.html?pageconfig=resource&rid=10372&pid=10000&source=pagetrap&viapagetrap=abpromise.

Respecto al secundario conjugado en verde, hay muchas alternativas disponibles en el catálogo. Puedes consultar las alternativas conjugadas en https://www.abcam.com/index.html?t=6013&pt=7&c=3050&ei=&mu=pt, https://www.abcam.com/index.html?t=6013&pt=7&c=5034&ei=&mu=pt o https://www.abcam.com/index.html?t=6013&pt=7&c=1874&ei=&mu=pt.

Si quisieras más información sobre alguna de ellas, no dudes en hacérmelo saber.

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1-10 of 22 Abreviews or Q&A

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