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Tags & Cell Markers Fusion / Marker Proteins GFP

Anti-GFP抗体(ab5450)

  • Datasheet
  • SDS
Reviews (10)Q&A (6)References (146)

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Abpromise

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-GFP antibody (ab5450)
  • Immunocytochemistry/ Immunofluorescence - Anti-GFP antibody (ab5450)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-GFP antibody (ab5450)
  • IHC - Wholemount - Anti-GFP antibody (ab5450)
  • Electron Microscopy - Anti-GFP antibody (ab5450)

Key features and details

  • Goat polyclonal to GFP
  • Suitable for: IP, ICC, IHC-FoFr, Electron Microscopy, IHC - Wholemount, ICC/IF, WB, IHC-P
  • Reacts with: Species independent
  • Isotype: IgG

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概述

  • 产品名称

    Anti-GFP抗体
    参阅全部 GFP 一抗
  • 描述

    山羊多克隆抗体to GFP
  • 宿主

    Goat
  • 经测试应用

    适用于: IP, ICC, IHC-FoFr, Electron Microscopy, IHC - Wholemount, ICC/IF, WB, IHC-Pmore details
  • 种属反应性

    与反应: Species independent
  • 免疫原

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

  • 阳性对照

    • Pure GFP protein, or cells known to overexpress GFP.
  • 常规说明

    Protein A will not bind goat IgG, so use alternates (eg. protein G) in IP with this antibody.

    Reproducibility is key to advancing scientific discovery and accelerating scientists’ next breakthrough.

    Abcam is leading the way with our range of recombinant antibodies, knockout-validated antibodies and knockout cell lines, all of which support improved reproducibility.

    We are also planning to innovate the way in which we present recommended applications and species on our product datasheets, so that only applications & species that have been tested in our own labs, our suppliers or by selected trusted collaborators are covered by our Abpromise™ guarantee.

    In preparation for this, we have started to update the applications & species that this product is Abpromise guaranteed for.

    We are also updating the applications & species that this product has been “predicted to work with,” however this information is not covered by our Abpromise guarantee.

    Applications & species from publications and Abreviews that have not been tested in our own labs or in those of our suppliers are not covered by the Abpromise guarantee.

    Please check that this product meets your needs before purchasing. If you have any questions, special requirements or concerns, please send us an inquiry and/or contact our Support team ahead of purchase. Recommended alternatives for this product can be found below, as well as customer reviews and Q&As.

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 or -80°C. Avoid freeze / thaw cycle.
  • 存储溶液

    pH: 7.40
    Constituents: 0.79% Tris HCl, 25% Glycerol
  • Concentration information loading...
  • 纯度

    Protein G purified
  • 纯化说明

    This 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.
  • 克隆

    多克隆
  • 同种型

    IgG
  • 研究领域

    • Tags & Cell Markers
    • Fusion / Marker Proteins
    • GFP

相关产品

  • Compatible Secondaries

    • Donkey Anti-Goat IgG H&L (Alexa Fluor® 488) (ab150129)
    • Donkey Anti-Goat IgG H&L (HRP) (ab205723)
  • Isotype control

    • Goat IgG, polyclonal - Isotype Control (ab37373)
  • Recombinant Protein

    • Recombinant E. coli GFP protein (His tag) (ab119740)
    • Recombinant A. victoria GFP protein (ab84191)
  • Related Products

    • Prestained Protein Ladder - Broad molecular weight (10-245 kDa) (ab116028)
    • GFP ELISA Kit (ab171581)
    • Anti-GFP antibody (ab5449)

应用

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.

应用 Ab评论 说明
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.

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

图片

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-GFP antibody (ab5450)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-GFP antibody (ab5450)Image from Sylva M et al., Plos One. 2011;6(10):e26088. Fig 2.; doi: 10.1371/journal.pone.0022616 Reproduced under the Creative Commons license http://creativecommons.org/licenses/by/4.0/

    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. 

  • Immunocytochemistry/ Immunofluorescence - Anti-GFP antibody (ab5450)
    Immunocytochemistry/ Immunofluorescence - Anti-GFP antibody (ab5450)

    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).

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-GFP antibody (ab5450)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-GFP antibody (ab5450)Image courtesy of an anonymous Abreview.
    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.
  • IHC - Wholemount - Anti-GFP antibody (ab5450)
    IHC - Wholemount - Anti-GFP antibody (ab5450)Image courtesy of Sana Zakaria by Abreview.
    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 - Anti-GFP antibody (ab5450)
    Electron Microscopy - Anti-GFP antibody (ab5450)This image is courtesy of an Abreview submitted by Narciso Campos

    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

实验方案

  • Immunoprecipitation protocols
  • Immunohistochemistry protocols
  • Immunocytochemistry & immunofluorescence protocols
  • Western blot protocols

Click here to view the general protocols

数据表及文件

    • Datasheet
    • SDS
  • 文献 (146)

    发表研究结果有使用 ab5450?请让我们知道,以便我们可以引用本数据表中的参考文章。

    ab5450 被引用在 146 文献中.

    • Zhou YQ  et al. Genetically engineered distal airway stem cell transplantation protects mice from pulmonary infection. EMBO Mol Med 12:e10233 (2020). PubMed: 31782624
    • Diao L  et al. Increased proliferation of hepatic periportal ductal progenitor cells contributes to persistent hypermetabolism after trauma. J Cell Mol Med 24:1578-1587 (2020). PubMed: 31793707
    • Costamagna D  et al. Interleukin-4 administration improves muscle function, adult myogenesis, and lifespan of colon carcinoma-bearing mice. J Cachexia Sarcopenia Muscle 11:783-801 (2020). PubMed: 32103619
    • Chillappagari S  et al. Defective BACH1/HO-1 regulatory circuits in cystic fibrosis bronchial epithelial cells. J Cyst Fibros N/A:N/A (2020). PubMed: 32534959
    • Eagle AL  et al. Circuit-specific hippocampal ?FosB underlies resilience to stress-induced social avoidance. Nat Commun 11:4484 (2020). PubMed: 32901027
    View all Publications for this product

    客户评价及客户问答

    Show All 评价 Q&A
    提交评价 提交问题

    1-10 of 16 Abreviews or Q&A

    Question

    We have 2 of your GFP antibodies
    Abcam # ab5450
    ab290
    I'd like to do IP and Western blot using these antibodies.
    My proteins are mTurquoise or mCitrine tagged.
    Do the above antibodies recognize mTurquoise or mCitrine?

    Read More

    Abcam community

    Verified customer

    Asked on Oct 16 2012

    Answer

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

    Read More

    Abcam Scientific Support

    回复于 Oct 16 2012

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) abreview for Anti-GFP antibody

    Excellent
    Abreviews
    Abreviews
    abreview image
    Application
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
    Sample
    Mouse Tissue sections (Uterus)
    Antigen retrieval step
    Heat mediated - Buffer/Enzyme Used: Sodium citrate
    Permeabilization
    Yes - Tween-20
    Specification
    Uterus
    Blocking step
    Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 23°C
    Fixative
    Paraformaldehyde
    Read More

    Reshef Tal

    Verified customer

    提交于 Sep 06 2019

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) abreview for Anti-GFP antibody

    Excellent
    Abreviews
    Abreviews
    abreview image
    Application
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
    Sample
    Rat Tissue sections (Skin)
    Antigen retrieval step
    Heat mediated - Buffer/Enzyme Used: Citrate
    Permeabilization
    No
    Specification
    Skin
    Blocking step
    Serum as blocking agent for 30 minute(s) · Concentration: 10% · Temperature: 22°C
    Fixative
    Formaldehyde
    Read More

    Abcam user community

    Verified customer

    提交于 Mar 18 2019

    IHC - Wholemount abreview for Anti-GFP antibody

    Excellent
    Abreviews
    Abreviews
    abreview image
    Application
    IHC - Wholemount
    Sample
    Mouse Tissue (trachea)
    Specification
    trachea
    Read More

    Abcam user community

    Verified customer

    提交于 Dec 11 2018

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) abreview for Anti-GFP antibody

    Good
    Abreviews
    Abreviews
    abreview image
    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
    Read More

    Abcam user community

    Verified customer

    提交于 Jun 18 2016

    Question

    Will this antibody react with mCherry or dsRed?

    Read More

    Abcam community

    Verified customer

    Asked on Oct 01 2015

    Answer

    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

    Read More

    Heather Allen

    Abcam Scientific Support

    回复于 Oct 01 2015

    Other (Immunolabeling for electron microscopy) abreview for Anti-GFP antibody

    Excellent
    Abreviews
    Abreviews
    abreview image
    Application
    Other - Do not use
    Sample
    Arabidopsis thaliana Tissue sections (True leaf from 8-day old seedling)
    Read More

    Mr. Narciso Campos

    Verified customer

    提交于 Jun 25 2013

    Question

    Dilucion recomendada para EM.

    Read More

    Abcam community

    Verified customer

    Asked on Mar 18 2013

    Answer

    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 muestras, pero en general, aconsejamos usar tres diluciones para empezar: 1/250 (1ug/250 ul), 1/1000 (1ug/1000ul) y 1/2500 (1ug/2.5ml).
    Si tienes cualquier otra consulta no dudes en volverme a contactar.

    Read More

    Abcam Scientific Support

    回复于 Mar 18 2013

    Question


    im Anhang finden sie zwei IHC-Figures und den ausgefüllten Fragebogen.
    Mit freundlichen Grüßen

    Read More

    Abcam community

    Verified customer

    Asked on Sep 05 2012

    Answer

    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 dies absolut möglich ist, auch aufgrund des hohen Titers dieses Antikörpers.

    Um auch allfällige technische Problem auszuschliessen, könnten folgende Punkte noch in Betracht gezogen werden, falls dies noch nicht gemacht wurde:

    1.) Haben Sie eine Isotypen Kontrolle gemacht, um allfällig unspezifische Färbung auszuschliessen?

    2.) Haben Sie eine Kontrolle in nicht-transgenem Gewebe gemacht? Dies würde erlauben, wieder Hintergrund und auch endogene CXCR4 Expression auszuschliessen.

    3.) Der UMB2 Antikörper erkennt ja nur ein Epitop auf dem CXCR4 Protein (im C Terminus). Vielleicht erkennt dieser Antikörper dann nur das Protein, wenn das Epitop freistehen ist- vielleicht, absolut hypothetisch, könnte dies anders sein einer neuenIsoform, modifizierter Form oder in einemProtein Complex, welche dannnicht in allen Zellen exprimiert wird. Vielleicht könnten Sie parallel zu dem UMB2 auch einen polyklonalen Antikörper gegen das ganze Protein von CXCR4 ausprobieren?

    Ich kenne leider keinen Antikörper gegen GFP/EGFP, welcher getestet wurde, dass er nur dasreife Protein erkennt.

    Bitte lassen Sie mich wissen, was Sie von den oben genannten Vorschlägen halten. Falls Sie einen anderen CXCR4 Antikörper testen wollen, können wir Ihnen vielleicht auch ein Testangebot machen.

    Ich würde mich freuen, wieder von Ihnen zu hören.

    Read More

    Abcam Scientific Support

    回复于 Sep 05 2012

    Question

    Customer kindly called to ask for any information which we may have regarding use of these products in EM.

    Read More

    Abcam community

    Verified customer

    Asked on May 23 2012

    Answer

    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.ncbi.nlm.nih.gov/pubmed/18187503?dopt=Abstract

    AB5450:

    -http://www.ncbi.nlm.nih.gov/pubmed/18385325?dopt=Abstract

    AB6556:

    -http://www.ncbi.nlm.nih.gov/pubmed/20140253?dopt=Abstract

    -http://www.ncbi.nlm.nih.gov/pubmed/19270696?dopt=Abstract

    Read More

    Abcam Scientific Support

    回复于 May 23 2012

    1-10 of 16 Abreviews or Q&A

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