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
  • Species reactivity
    Reacts with: Species independent
  • Immunogen

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

  • Positive control
    • IHC: E5.5 Hex-GFP transgenic mouse embryo. WB: HeLa cells. Green Fluorescent protein.

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

  • Pth4:eGFP transgenic zebrafish embryos at 1 and 2 dpf were fixed with 4% PFA and washed in PBST. They were then washed in PBDT (1% BSA, 1% DMSO, 0.1% Triton X-100 in PBS, pH 7.4), blocked in 10% normal goat serum/PBDT, and incubated overnight at 4°C with primary antibodies to HuC/D (1/100) and GFP (1/400, Abcam ab6673). Further PBST washes and blocking were followed by secondary antibodies overnight at 4°C. Hoechst 34580 was added to stain nuclei (1/2500). After further PBDT and PBS washes, embryos were mounted for confocal imaging.

    Abbreviation: e, eye; hy, hypothalamus; m, midbrain; sc, spinal cord. Scale bars: 100 μm (A-C) 50 μm (D-G).

  • In utero electroporation of Disc1 and Disc1-100P constructs into wild-type neocortex and analysis at P21.

    (Panels D-E”) Expression of the constructs was assessed.

    (Panels D-D'') 2 days after transfection in vitro.

    (Panels E-E'') at P21 in vivo.

    Immunochemistry for FLAG and GFP showed that constructs encoding either WT Disc1, the Disc1-100P variant, or GFP alone, expressed these protein species in transfected HEK-293 cells in vitro (Fig 5D–5D”) and in P21 postmitotic cortical neurons in vivo (Fig 5E–5E”)

  • Immunofluorescence for assessment of GFP+ myofibers in rat tissue.

    VML affected muscle from the 50% MG + HA+LMN group were probed for the presence of GFP. GFP+ fibers were detected in a qualitatively similar magnitude at both 2 and 8 weeks post-injury indicating viable engraftment of donor derived muscle progenitor cells. Scale bars are 1mm for whole mount images, 50 μm for regions of interest.

    A portion of the TA muscle from the defect region was embedded in a talcum-based gel, frozen in 2-methylbutane, and supercooled in liquid nitrogen. Cryosections (8 μm) were prepared and stained using standard protocols for hematoxylin & eosin.

    ab6673 used at a 1/100 dilution.

  • Mouse small intestines were washed with DPBS and fixed overnight at 4°C in Zinc formalin. Following sectioning and tissue deparaffanization, antigen retrieval was performed with 10mM Tris base (pH 9.0) buffer using a pressure cooker.

    For immunohistochemistry, sections were quenched of endogenous peroxidases by 3% H2O2, and sequentially blocked with Avidin D, biotin, and protein blocking reagents. Primary antibody incubation was conducted at 4°C overnight. Secondary biotinylated antibody was added at a dilution of 1/200, and incubated 2 hours at room temperature. Finally, sections were stained according to the ABC peroxidase protocol and counterstained with hematoxylin.

    ab6673 used at a 1/200 dilution.

    Panel D: Representative anti-eGFP immunofluorescence of macroH2A WT and DKO jejunum counterstained with DAPI (blue).

  • All lanes : Anti-GFP antibody (ab6673) at 1 µg/ml (o/n at 4degC)

    Lane 1 : HEK-293 (Human epithelial cell line from embryonic kidney) lysate at 10 µg
    Lane 2 : HeLa (Human epithelial cell line from cervix adenocarcinoma) lysate at 10 µg
    Lane 3 : CHO/K1 lysate at 10 µg
    Lane 4 : MDA-MB-231 (Human breast adenocarcinoma cell line) lysate at 10 µg
    Lane 5 : A431 (Human epidermoid carcinoma cell line) lysate at 10 µg
    Lane 6 : Jurkat (Human T cell leukemia cell line from peripheral blood) lysate at 10 µg
    Lane 7 : NIH/3T3 (Mouse embryo fibroblast cell line) lysate
    Lane 8 : E-coli HCP control, 50 ng
    Lane 9 : FLAG Positive control lysate at 10 µg
    Lane 10 : Red fluorescent protein, 50 ng
    Lane 11 : Green fluorescent protein, 50 ng
    Lane 12 : Glutathinoe-S-Transferase protein, 50 ng
    Lane 13 : Maltose Binding protein, 50 ng

    Secondary
    All lanes : Peroxidase goat secondary antibody, 60 min at RT at 1/30000 dilution


    Blocking Buffer: 1% Casein-TTBS for 30 min at RT.

  • E5.5 Hex-GFP transgenic mouse embryo stained for GFP using ab6673 at 1/500 dilution. Secondary antibody is a fluorochrome conjugated anti-goat IgG secondary antibody at 1/10,000 for 45 min at RT.

    Staining: GFP as green fluorescent signal with DAPI blue counterstain.

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

    Lane 1 : HeLa (Human epithelial cell line from cervix adenocarcinoma) cells
    Lane 2 : Mock transfected HeLa cell lysate

    Lysates/proteins at 35 µg per lane.

    Secondary
    All lanes : IRDye® 800 conjugated Donkey-a-Goat IgG [H&L] at 1/2500 dilution

    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

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

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

References

This product has been referenced in:
See all 212 Publications for this product

Customer reviews and Q&As

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1-10 of 16 Abreviews

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

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
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

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

Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Mouse Tissue sections (pancreas)
Specification
pancreas
Fixative
Formaldehyde
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: citrate, ph6
Permeabilization
No
Blocking step
(agent) for 10 minute(s) · Concentration: 100% · Temperature: RT°C

Dr. Miri Stolovich-Rain

Verified customer

Submitted Apr 03 2012

Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Human Tissue sections (Liver)
Specification
Liver
Fixative
Paraformaldehyde
Antigen retrieval step
None
Permeabilization
No
Blocking step
Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 10% · Temperature: 20°C

Abcam user community

Verified customer

Submitted May 17 2011

1-10 of 16 Abreviews

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