Overview

  • Product name
  • Description
    Chicken polyclonal to GFP
  • Host species
    Chicken
  • Tested applications
    Suitable for: IHC-P, WB, IHC - Wholemount, IHC-FrFl, ICC/IF, IHC-Fr, IHC-FoFrmore details
  • Immunogen

    Recombinant full length protein corresponding to GFP.
    Database link: P42212

  • Positive control
    • WB: Transgenic mouse spinal cords. ICC/IF: GFP-transfected NIH/3T3 cells.

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
    Preservative: 0.01% Thimerosal (merthiolate)
    Constituents: PBS, 50% Glycerol, 0.16% Sodium phosphate
  • Concentration information loading...
  • Purity
    IgY fraction
  • Purification notes
    Sterile filtered.
  • Clonality
    Polyclonal
  • Isotype
    IgY
  • Research areas

Applications

Our Abpromise guarantee covers the use of ab13970 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
IHC-P 1/500 - 1/1000. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

The concentrations of fixative for the IHC applications were typically 10% formalin or 2% paraformaldehyde.

WB 1/5000.
IHC - Wholemount Use at an assay dependent concentration.
IHC-FrFl Use at an assay dependent concentration.
ICC/IF 1/2000.

Used at a dilution of 1/2000 for 1 hr (see Abreview for further information).

IHC-Fr 1/1000.
IHC-FoFr 1/2000.

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

  • Adenoviral Cre induces mosaic activation of rtTA and GFP induction in CAGs-LSL-rtTA3 and CAGs-LSL-RIK animals.

    Immunofluorescent stains for GFP and mKate2 in lung sections of triple transgenic mice (CAGs-LSL-rtTA3 or RIK;TG-Ren.713;LSL-KrasG12D). KrasG12D-induced lung adenomas show strong expression of GFP and mKate2.

    Lower panel: Higher magnification of the lesion.

    White arrows indicate rare cells that show mKate2, but not GFP expression.

  • All lanes : Anti-GFP antibody (ab13970) at 1/200 dilution

    Lane 1 : Wild type 'naive' HEK293 whole cell lysates
    Lanes 2-4 : GFP transfected HEK293 whole cell lysates


    Lysates/proteins at 5 µg per lane.

    Secondary
    All lanes : Donkey anti-chicken polyclonal CY5 conjugate. at 1/1000 dilution

    Performed under reducing conditions.

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


    Exposure time: 25 minutes


    Cells were treated by mixing in RIPA buffer and denaturating in Laemmli buffer for 5 minutes at 95°C. The gel was Precast at 4-12%. Blocking was with 0.5% milk at 20°C for 5 minutes.

    See Abreview

  • ab13970 staining mouse olfactory bulb tissue sections by IHC-Fr.

    Sections were PFA fixed, permeabilized in 0.4% Triton-X and blocked with 5% serum for 2 hours at 25°C.  The primary antibody was diluted 1/1000 and incubated with the sample for 16 hours at 4°C.  An Alexa Fluor® 488 conjugated goat anti-chicken was used as the secondary.

    See Abreview

  • ab13970 staining GFP in murine lung tissue by Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections).

    Tissue was fixed in paraformaldehyde and a heat mediated antigen retrieval step was performed using citrate buffer. Samples were then permeabilized with 0.1% Tween, blocked with 15% serum for 30 minutes at 23°C and then incubated with ab13970 at a 1/500 dilution for 14 hours at 4 °C. The secondary used was an Alexa-Fluor® 488 conjugated goat anti-chicken polyclonal used at a 1/500 dilution.

    See Abreview

  • ab13970 staining GFP in murine olfactory bulb tissue by Immunohistochemistry (PFA perfusion fixed frozen sections) Counterstained with DAPI.
  • ab13970 staining GFP in U-2 OS (Human bone osteosarcoma epithelial cell line) cells by ICC/IF.

    Cells were paraformaldehyde fixed, permeabilized with 0.5% triton and blocked with 2% antibody dilution buffer for 2 hours. Cells were incubated with the primary antibody (1/1000) for 1 hour at 25°C. An undiluted Alexa Fluor® 488 conjugated Goat anti-chicken polyclonal was used as the secondary antibody.

    See Abreview

  • All lanes : Anti-GFP antibody (ab13970) at 1/1000 dilution

    Lane 1 : Whole cell lysate prepared from HeLa (Human epithelial cell line from cervix adenocarcinoma) cells.
    Lane 2 : Whole cell lysate prepared from HeLa cells.

    Lysates/proteins at 25 µg per lane.

    Secondary
    All lanes : IRDye 800CW conjugated goat anti-chicken polyclonal at 1/15000 dilution

    See Abreview

  • Transgenic mice expressing GFP selectively in lamina II of the spinal cord.

    In the right panels, note the correspondance between the green (rabbit anti-GFP) and red signals (chicken anti-GFP from Abcam) indicating that these two antibody preparations recognized the same gene product. The secondary antibody used with ab13970 was an FITC-labeled goat anti-chicken

  • ab13970 staining GFP in GFP-transfected NIH/3T3 (Mouse embryo fibroblast cell line) cells.

    The cells were fixed with 4% formaldehyde (10 minutes) and then blocked in 1% BSA / 0.3M glycine in 0.1%PBS-Tween for 1 hour. The cells were then incubated with ab13970 at 1/2000 dilution overnight at +4°C followed by incubation with Goat Anti-Chicken IgY H&L (Alexa Fluor® 488) preadsorbed (ab150173), for 1 hour, at 1 μg/ml.
    Under identical experimental conditions, when compared to the basal level of GFP expression in transfected NIH/3T3 cells, the cells upon which ab13970 was applied gave a stronger signal in the 488 channel, indicating that ab13970 is binding to GFP and therefore eliciting signal amplification.
    ab13970 was also applied to non-GFP-transfected NIH/3T3 cells, which produced no positive staining, indicating specificity for GFP.

    Nuclear DNA was labeled with 1.43 μM DAPI (blue).

  • Lane 1 : Rabbit anti-GFP
    Lane 2 : Anti-GFP antibody (ab13970)

    All lanes : Transgenic mouse spinal cords


    Western blot of transgenic mouse spinal cords showing that the rabbit anti-GFP (lane 1) and the chicken anti-GFP (Abcam; lane 2) recognize a band at the same molecular weight.

  • ab13970 staining mouse olfactory epithelium tissue sections by IHC-Fr.

    The sample was PFA fixed and blocked with 4% BSA/ 5% NFDM/ 10% NDS for 15 minutes at 20°C. The primary antibody was diluted 1/1000 and incubated with the sample for 1 hour.  An FITC conjugated goat anti-chicken was used as the secondary.

    This colony is the result of retroviral infection with a control virus. The GFP is under the control of an IRES promoter, so its expression is independent of any other protein. The counterstain is hoescht.

    See Abreview

  • ab13970 staining GFP + tumor in mouse muscle cells by ICC/IF.

    Cells were formaldehyde fixed and blocked with 3% BSA for 1 hour at 24°C prior to incubation with the primary antibody (1/500) for 1 hour at 24°C. An Alexa Fluor® 488 conjugated goat anti-chicken was used as the secondary.

    See Abreview

  • ab13970 staining GFP in mouse brain tissue section by Immunohistochemistry (PFA perfusion fixed frozen sections).

    Tissue samples were fixed with paraformaldehyde and permeabilized with 0.1% Triton X100 before blocking with 10% serum for 30 minutes at 25°C. The sample was incubated with primary antibody (1/2000) for 16 hours at 25°C in 10% NGS in PBS + 0.1% TX100. An Alexa Fluor®488-conjugated Goat polyclonal to chicken IgG was used as secondary antibody at 1/400 dilution.

    In the image, green staining represents GFP expressed in oligodendrocytes, blue is for ToPro3.

    See Abreview

  • Immunocytochemical/ Immunoflurescence analysis of cytospined HEK-293 cells (Human epithelial cell line from embryonic kidney) transfected with GFP, labeling GFP with ab13970 at 1/200 incubated for 16 hours at 4°C with 1% BSA in PBS. Secondary used was a donkey anti-chicken polyclonal DyLight® 594 at 1/500.

    GFP is shown in red (DyLight® 594).

    Nuclei are counterstained in blue (DAPI).

    The left panel shows HEK-293 cells transfected with GFP and the right panel shows non-transfected HEK-293 cells.

    See Abreview

References

This product has been referenced in:
  • Aguillon R  et al. Cell-type heterogeneity in the early zebrafish olfactory epithelium is generated from progenitors within preplacodal ectoderm. Elife 7:N/A (2018). Read more (PubMed: 29292696) »
  • White MG  et al. Anterior Cingulate Cortex Input to the Claustrum Is Required for Top-Down Action Control. Cell Rep 22:84-95 (2018). Read more (PubMed: 29298436) »
See all 1085 Publications for this product

Customer reviews and Q&As

1-10 of 104 Abreviews or Q&A

Application
IHC - Wholemount
Sample
Fruit fly (Drosophila melanogaster) Tissue (Adult Posterior Midgut, HEAT-FIXED)
Specification
Adult Posterior Midgut, HEAT-FIXED

Dr. Joaquin Denavascues

Verified customer

Submitted Feb 12 2014

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunocytochemistry/ Immunofluorescence
Blocking step
BSA as blocking agent for 40 minute(s) · Concentration: 5% · Temperature: 22°C
Sample
Human Cell (HEK293)
Specification
HEK293
Permeabilization
Yes - 0.2% triton
Fixative
Formaldehyde

Abcam user community

Verified customer

Submitted Sep 26 2013

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
Sample
Human Cell lysate - whole cell (HEK 293)
Specification
HEK 293
Treatment
transfection with 1 ug GFP plasmid
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C

Abcam user community

Verified customer

Submitted Aug 01 2013

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunocytochemistry/ Immunofluorescence
Blocking step
Abdil buffer as blocking agent for 2 hour(s) and 0 minute(s) · Concentration: 2%
Sample
Human Cell (U2OS)
Specification
U2OS
Permeabilization
Yes - 0.5% triton
Fixative
Paraformaldehyde

Dr. Christophe Lachaud

Verified customer

Submitted Aug 01 2013

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunocytochemistry/ Immunofluorescence
Blocking step
Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 22°C
Sample
Human Cell (U2OS cells)
Specification
U2OS cells
Permeabilization
Yes - 0.2% Triton
Fixative
Paraformaldehyde

Abcam user community

Verified customer

Submitted Jul 26 2013

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

Dr. Francesco Elia Marino

Verified customer

Submitted Sep 26 2018

Application
Immunohistochemistry free floating
Sample
Mouse Tissue sections (nerve)
Specification
nerve

Abcam user community

Verified customer

Submitted Aug 31 2018

Application
Immunohistochemistry (Frozen sections)
Sample
Mouse Tissue sections (Brain)
Permeabilization
Yes - 0.1% Triton X-100
Specification
Brain
Blocking step
Serum as blocking agent for 24 hour(s) and 0 minute(s) · Concentration: 10% · Temperature: 4°C
Fixative
Paraformaldehyde

Kevin Zhang

Verified customer

Submitted Jan 31 2018

Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Mouse Tissue sections (Prostate)
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: Citrate, pH 6
Permeabilization
No
Specification
Prostate
Blocking step
Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 10% · Temperature: 20°C
Fixative
Paraformaldehyde

Abcam user community

Verified customer

Submitted Dec 12 2017

Application
IHC - Wholemount
Sample
Human Embryo (H9-gfp cells)
Specification
H9-gfp cells

Abcam user community

Verified customer

Submitted Apr 12 2017

1-10 of 104 Abreviews or Q&A

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

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