Overview

  • Product nameAnti-GFP antibody
    See all GFP primary antibodies
  • Description
    Goat polyclonal to GFP
  • Tested applicationsSuitable for: IP, ICC, IHC-FoFr, Electron Microscopy, IHC - Wholemount, ICC/IF, WB, IHC-Pmore details
  • Immunogen

    Recombinant full length protein corresponding to GFP.

  • Positive control
    • GFP-transfected NIH3T3 cells
  • General notesProtein A will not bind goat IgG, so use alternates (eg. protein G) in IP with this antibody.

Properties

  • FormLiquid
  • Storage instructionsShipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C or -80°C. Avoid freeze / thaw cycle.
  • Storage bufferPreservative: None
    Constituents: 25% Glycerol, 50mM Tris HCl. pH 7.4
  • Concentration information loading...
  • PurityProtein G purified
  • Purification notesThis antibody is Goat anti-GFP serum (ab5449) affinity purified using a HiTrap-NHS activated sepharose column (Amersham-Pharmacia) containing covalently linked highly purified recombinant GFP. After applying the serum to the column and extensive washing, the eluted anti-GFP immunoglobulins are desalted.
  • ClonalityPolyclonal
  • IsotypeIgG
  • Research areas

Associated products

Applications

Our Abpromise guarantee covers the use of ab5450 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
IP Use at an assay dependent concentration.
ICC Use at an assay dependent concentration.
IHC-FoFr Use at an assay dependent concentration.
Electron Microscopy Use at an assay dependent concentration.
IHC - Wholemount Use at an assay dependent concentration.
ICC/IF 1/2000.
WB 1/2000 - 1/20000.
IHC-P 1/1000.

Target

  • RelevanceFunction: 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
    • yfp antibody

Anti-GFP antibody images

  • Immunohistochemical staining of donor BMDCs expressing GFP in mouse pancreas tissue using ab5450. Antigen retrieval was carried out using target-retrieval solution for 30 minutes in a boiling water bath. Endogenous peroxidase activity was quenched with 3% hydrogen peroxide (5 minutes). Panels A and B were detected with a polymer horseradish peroxidase anti-rabbit detection system (30 minutes) and 3,3′-diaminobenzidine was used as a substrate. Counter-staining was performed with hematoxylin. 

  • ab5450 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 ab5450 at 1/2000 dilution overnight at +4°C followed by incubation with ab150129, Donkey Anti-Goat 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 ab5450 was applied gave a stronger signal in the 488 channel, indicating that ab5450 is binding to GFP and therefore eliciting signal amplification.
    ab5450 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).

  • ab5450 staining GFP in murine brain tissue by Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections). Tissue was fixed with paraformaldehyde, permeabilized using 0.2% Triton and then blocked with serum for 1 hour at room temperature, followed by incubation with the primary antibody at a 1/1000 dilution for 14 hours at 4°C. A Cy2®-conjugated secondary antibody was used at a 1/200 dilution.
  • ab5450 staining GFP in murine hindbrain tissue by IHC-Wholemount.
    Dissected hindbrains of E12.5 stage embryos were fixed in 4% paraformaldehyde for 4-5 hours and then washed in PBS before proceeding with immunolabelling in PBS with 1% triton. Primary antibody was incubated for 4-6 days (99-144 hours). The hindbrain was cultured as an explant after GFP was electroporated. The wholemount immuno with ab5450 at a 1/500 dilution was done to see if GFP could be detected, it was the same GFP expression with the antibody as that which had been seen initially with the electroporation. The secondary used was an Alexa-Fluor 647 conjugated donkey anti-goat polyclonal used at a 1/150 dilution. Blue is DAPI and green is the GFP antibody (pseudo-colored from red Alexa Fluor 647).

    See Abreview

  • Electron Microscopy of Arabidopsis thaliana tissue sections labelling GFP with ab5450. An 18nm gold-conjugated Donkey anti-goat IgG polyclonal (1/15) was used as the secondary antibody.

     

    Arabidopsis thaliana transgenic plant expressing GFP fused to an Endoplasmic Recticulum (ER) marker. No label is observed in: C - chloroplasts, M - mitochondria, G - golgi. The sample was prepared by cryofixation and embedding in Lowicryl HM20 resin. The image was taken in a JEOL 1010 transmission electron microscope at 20,000× magnification.

    See Abreview

References for Anti-GFP antibody (ab5450)

This product has been referenced in:
  • Drelon C  et al. PKA inhibits WNT signalling in adrenal cortex zonation and prevents malignant tumour development. Nat Commun 7:12751 (2016). IHC-P . Read more (PubMed: 27624192) »
  • Wigington CP  et al. Post-transcriptional regulation of programmed cell death 4 (PDCD4) mRNA by the RNA-binding proteins human antigen R (HuR) and T-cell intracellular antigen 1 (TIA1). J Biol Chem 290:3468-87 (2015). ICC/IF . Read more (PubMed: 25519906) »

See all 45 Publications for this product

Product Wall

Since mTurquoise or mCitrine originate from Aquorea victoria jelly fish, then they will react with these GFP antibodies.

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, polymorphic layer of the dentate gyrus)
Antigen retrieval step Heat mediated - Buffer/Enzyme Used: citrate buffer pH6
Specification brain, polymorphic layer of the dentate gyrus
Blocking step BSA as blocking agent for 40 minute(s) · Concentration: 1% · Temperature: RT°C
Fixative Paraformaldehyde
Username

Abcam user community

Verified customer

Submitted Jun 18 2016

It is highly likely that they will react with mCherry although this hasn't been specifically tested. However, it will react with dsRed that is very similar to mCherry

Application Other
Sample Arabidopsis thaliana Tissue sections (True leaf from 8-day old seedling)
Username

Mr. Narciso Campos

Verified customer

Submitted Jun 25 2013

Te pido disculpas por la espera.

En relación a la dilución recomendada para usar el anticuerpo en Microscopia Electrónica, lógicamente dependerá del nivel de expresión de la proteína en las mue...

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Vielen Dank für das Zusenden der detaillierten Daten. Ich entschuldige mich für die lange Zeit, welche Sie auf eine Antwort warten mussten.

Das Labor hat leider nicht getested ob der ab5450 die Pre-Form erkennt, denkt jedoch, dass d...

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All three products which we spoke of today have been tested in EM with publication. I have included links to those for your review below:

AB290:

-http://www.ncbi.nlm.nih.gov/pubmed/18574249?dopt=Abstract

-http://www.ncb...

Read More
Abcam guarantees this product to work in the species/application used in this Abreview.
Application IHC - Wholemount
Sample Mouse Tissue (Hindbrain)
Specification Hindbrain
Username

Ms. Sana Zakaria

Verified customer

Submitted Mar 23 2012

Abreviews
Application ChIP
Sample Human Cell lysate - whole cell (HEK 293T cells)
Specification HEK 293T cells
Type Cross-linking (X-ChIP)
Duration of cross-linking step: 10 minute(s) and 0 second(s)
Specification of the cross-linking agent: as per ab500 ChIP kit
Detection step Semiquantitative PCR
Positive control used a construct that contained my promoter of interest as a positive control
Negative control pcr with primers that amplified the target gene and not the promoter
Username

Dr. Jonathan Rud

Verified customer

Submitted Jan 11 2010

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Immunoprecipitation
Sample Human Cell lysate - whole cell (HEK 293T cells)
Total protein in input 25 µg
Specification HEK 293T cells
Treatment Transfected with GFP tagged protein
Immuno-precipitation step Protein A/G
Username

Dr. Jonathan Rud

Verified customer

Submitted Jan 11 2010

1-10 of 13 Abreviews or Q&A

Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"