概述

  • 产品名称Anti-GFP抗体
    参阅全部 GFP 一抗
  • 描述
    鸡多克隆抗体to GFP
  • 经测试应用适用于: IHC-P, WB, IHC - Wholemount, IHC-FrFl, ICC/IF, IHC-Fr, IHC-FoFrmore details
  • 免疫原

    Recombinant full length protein corresponding to GFP.

  • 阳性对照
    • ICC/IF: GFP-transfected NIH3T3 cells

性能

  • 形式Liquid
  • 存放说明Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C. Avoid freeze / thaw cycle.
  • 存储溶液Preservative: 0.01% Thimerosal (merthiolate)
    Constituents: PBS, 50% Glycerol, 0.16% Sodium phosphate
  • Concentration information loading...
  • 纯度IgY fraction
  • 纯化说明Serile filtered.
  • 克隆多克隆
  • 同种型IgY
  • 研究领域

应用

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.

应用 Ab评论 说明
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.

靶标

  • 相关性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.
  • 别名
    • GFP antibody
    • Green fluorescent protein antibody
    • yfp antibody

Anti-GFP antibody 图像

  • 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

  • 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.
  • 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

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

    Lane 1 : Wild type 'naive' HEK293 whole cell lysates
    Lane 2 : GFP transfected HEK293 whole cell lysates
    Lane 3 : GFP transfected HEK293 whole cell lysates
    Lane 4 : GFP transfected HEK293 whole cell lysates
    Lane 5 : As above

    Lysates/proteins at 5 µg per lane.

    Secondary
    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

    Image is courtesy of an AbReview submitted by Dr Francois Daubeuf.

    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

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

    Lane 1 : Whole cell lysate prepared from HeLa cells.
    Lane 2 : Whole cell lysate prepared from HeLa cells.

    Lysates/proteins at 25 µg per lane.

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

    Image courtesy of an anonymous Abreview.

    See Abreview

  • 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

  • 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 ab150173, Goat polyclonal Secondary Antibody to Chicken IgY - 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 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).

  • 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 + 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 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 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

Anti-GFP antibody (ab13970)参考文献

This product has been referenced in:
  • Jara JH  et al. Healthy and diseased corticospinal motor neurons are selectively transduced upon direct AAV2-2 injection into the motor cortex. Gene Ther N/A:N/A (2016). ICC/IF ; Mouse . Read more (PubMed: 26704722) »
  • Sanges D  et al. Reprogramming Müller glia via in vivo cell fusion regenerates murine photoreceptors. J Clin Invest 126:3104-3116 (2016). IHC-P . Read more (PubMed: 27427986) »

See all 563 Publications for this product

Product Wall

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

提交于 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

提交于 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

提交于 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

提交于 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

提交于 Jul 26 2013

Application Immunocytochemistry/ Immunofluorescence
Sample Human Cell (U2OS)
Permeabilization Yes - triton 0.5%
Specification U2OS
Fixative Paraformaldehyde
Username

Abcam user community

Verified customer

提交于 Nov 11 2016

Application Immunocytochemistry/ Immunofluorescence
Sample Mouse Cell (Cultured primary cells (adherent))
Permeabilization Yes - 0.1% triton
Specification Cultured primary cells (adherent)
Blocking step Normal goat serum as blocking agent for 30 minute(s) · Concentration: 7% · Temperature: 25°C
Fixative Paraformaldehyde
Username

Abcam user community

Verified customer

提交于 Sep 09 2016

Application IHC - Wholemount
Sample Astatotilapia burtoni Tissue (Brain (CLARITY cleared with Passive clearing))
Specification Brain (CLARITY cleared with Passive clearing)
Username

Dr. Sebastian Alvarado

Verified customer

提交于 Aug 17 2016

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Immunocytochemistry/ Immunofluorescence
Sample Mouse Cell (myoblast)
Permeabilization Yes - 0.5% NP-40
Specification myoblast
Blocking step BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 3% · Temperature: 18°C
Fixative Formaldehyde
Username

Miss. Can Zhou

Verified customer

提交于 Mar 11 2016

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 (esophagus)
Antigen retrieval step Heat mediated - Buffer/Enzyme Used: citrate, pH6.0
Permeabilization No
Specification esophagus
Blocking step (agent) for 20 minute(s) · Concentration: 100% · Temperature: 20°C
Fixative Formaldehyde
Username

Ms. Tatiana Karakasheva

Verified customer

提交于 Mar 08 2016

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