製品の概要

  • 製品名
  • 製品の詳細
    Chicken polyclonal to GFP
  • 由来種
    Chicken
  • アプリケーション
    適用あり: IHC-P, WB, IHC - Wholemount, IHC-FrFl, ICC/IF, IHC-Fr, IHC-FoFrmore details
  • 免疫原

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

  • ポジティブ・コントロール
    • ICC/IF: GFP-transfected NIH3T3 cells

法規制情報

製品の特性

  • 製品の状態
    Liquid
  • 保存方法
    Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C. Avoid freeze / thaw cycle.
  • バッファー
    Preservative: 0.01% Thimerosal (merthiolate)
    Constituents: PBS, 50% Glycerol, 0.16% Sodium phosphate
  • Concentration information loading...
  • 精製度
    IgY fraction
  • 特記事項(精製)
    Sterile filtered.
  • ポリ/モノ
    ポリクローナル
  • アイソタイプ
    IgY
  • 研究分野

アプリケーション

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.

アプリケーション Abreviews 特記事項
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.

ターゲット情報

  • 関連性
    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.
  • 別名
    • GFP antibody
    • Green fluorescent protein antibody

画像

  • ab13970 staining GFP in Human U2OS 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 1/200 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


    Western blot analysis of HEK293 transfected and untransfected cell lysates, labelling GFP with ab13970. Cells were treated by mixing in RIPA buffer and denaturating in Laemmli buffer for 5 mins at 95°C. The gel was Precast at 4-12%. Blocking was with 0.5% milk at 20°C for 5 mins.

    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 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

  • 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.

    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.
  • 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 a FITC-labeled goat anti-chicken

  • ab13970 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 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 1μg/ml.
    Under identical experimental conditions, when compared to the basal level of GFP expression in transfected NIH3T3 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 NIH3T3 cells, which produced no positive staining, indicating specificity for GFP. Nuclear DNA was labelled with 1.43μM DAPI (blue).

  • 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.   A 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 counter-stain is hoescht.

    See Abreview

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

    All lanes : 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

  • 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 250C. The sample was incubated with primary antibody (1/2000) for 16 hours at 250C 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

  • ab13970 staining GFP in murine olfactory bulb tissue by Immunohistochemistry (PFA perfusion fixed frozen sections) Counterstained with DAPI.
  • Immunocytochemical immunoflurescence analysis of human cytospined HEK293 cells transfected with GFP, labelling 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 pane shows HEK293 cells transfected with GFP and the right pane shows non-transfected HEK293 cells.

    See Abreview

参考文献

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 929 Publications for this product

レビューと Q&A

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

Dr. Joaquin Denavascues

Verified customer

投稿 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
Username

Abcam user community

Verified customer

投稿 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
Username

Abcam user community

Verified customer

投稿 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
Username

Dr. Christophe Lachaud

Verified customer

投稿 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
Username

Abcam user community

Verified customer

投稿 Jul 26 2013

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
Username

Kevin Zhang

Verified customer

投稿 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
Username

Abcam user community

Verified customer

投稿 Dec 12 2017

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

Abcam user community

Verified customer

投稿 Apr 12 2017

Application
Immunohistochemistry (Frozen sections)
Sample
Mouse Tissue sections (Embryonic Heart)
Permeabilization
Yes - Tween-20
Specification
Embryonic Heart
Blocking step
Serum as blocking agent for 2 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 20°C
Fixative
Paraformaldehyde
Username

Abcam user community

Verified customer

投稿 Apr 06 2017

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunocytochemistry
Sample
Rat Cultured Cells (genetically modified fibroblasts)
Permeabilization
Yes - Tx-100
Specification
genetically modified fibroblasts
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 24°C
Fixative
Paraformaldehyde
Username

Mr. Tamas Bellak

Verified customer

投稿 Jan 26 2017

1-10 of 102 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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