製品の概要

  • 製品名Anti-GFP antibody
    GFP 一次抗体 製品一覧
  • 製品の詳細
    Rabbit polyclonal to GFP
  • 特異性This antibody is reactive against all variants of Aequorea victoria GFP such as S65T-GFP, RS-GFP, YFP, CFP, RFP and EGFP.
  • アプリケーション適用あり: IHC-FoFr, Electron Microscopy, IHC-P, IHC-Fr, IP, WB, ICC/IF, Flow Cytmore details
  • 免疫原

    Recombinant full length protein corresponding to GFP.

  • ポジティブ・コントロール
    • Pure GFP protein, or cells known to overexpress GFP.
  • 特記事項

    Please note that a mistake was made in reference 4 (Mesaeli et.al., J. Cell. Biol. 1999 Mar 8;144(5):857-68). The antibody used for immunohistochemistry on paraformaldehyde fixed tissues was the crude serum version of this antibody (Abcam ab290) and not Clontech's monoclonal as stated. This product is supplied in 25% glycerol. During freezeing and thawing some phase separation might occur - Please ensure that the solution is mixed thoroughly but GENTLY before use.

    This antibody (ab6556) is the purifed version of our best-selling rabbit polyclonal to GFP (ab290). It has been developed specifically for use in applications requiring a high titre and specificity with minimum background such as immuno-electron microscopy.

    This anti-GFP antibody recognizes the enhanced form of GFP as well.

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 or -80°C. Avoid freeze / thaw cycle.
  • バッファーPreservative: None
    Constituents: 25% Glycerol, 50mM Tris HCl. pH 7.4
  • Concentration information loading...
  • 精製度Protein A purified
  • 特記事項(精製)This antibody is an affinity purified rabbit anti-GFP antibody purified on an affinity chromatography column made with highly purified recombinant GFP.
  • ポリ/モノポリクローナル
  • アイソタイプIgG
  • 研究分野

アプリケーション

Our Abpromise guarantee covers the use of ab6556 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-FoFr Use at an assay dependent concentration. PubMed: 19563657
Electron Microscopy 1/5000.
IHC-P 1/1000. Perform heat mediated antigen retrieval via the microwave method before commencing with IHC staining protocol.
IHC-Fr 1/1000.
IP Use at an assay dependent concentration. Use Protein A agarose for IP.
WB 1/1000 - 1/5000.
ICC/IF 1/2000.
Flow Cyt Use at an assay dependent concentration.

ab171870 - Rabbit polyclonal IgG, is suitable for use as an isotype control with this antibody.

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
    • yfp antibody

Anti-GFP antibody 画像

  • All lanes : Anti-GFP antibody (ab6556) at 1/5000 dilution

    Lane 1 : LNCaP cells pEGFP-PKD1 empty vector
    Lane 2 : LNCaP pEGFP-PKD1 transfected cells

    Secondary
    Rabbit polyclonal to GFP (ab6556) at 1/10000 dilution (Goat anti-rabbit HRP)
    Developed using the ECL technique

    Performed under reducing conditions.

    Exposure time : 10 seconds

    See Abreview

  • ab6556 at 1/500 dilution staining GFP in mouse testis by immunohistochemistry (frozen sections). Sections were paraformaldehyde fixed prior to blocking in 100% serum for 1 hour at 37°C and then incubated with ab6556 for 1 hour at 37°C. A Texas Red conjugated chicken polyclonal to rabbit Ig, diluted 1/100, was used as the secondary antibody.

    See Abreview

  • This image shows a single primary hippocampal neuron from a primary culture overexpressing GFP stained with ab6556 at a dilution of 1/2000. This picture was kindly supplied as part of the review submitted by one of our customers.

  • ab6556 staining GFP in mouse tooth tissue section by Immunohistochemistry (Frozen sections) without fixation. Tissue samples were blocked with 1% BSA for 20 minutes at 20oC. The sample was incubated with primary antibody (1/500) for 2 hours at 20oC. An Alexa Fluor®488-conjugated Chicken polyclonal to rabbit IgG was used as secondary antibody at 1/200 dilution. Immunofluorescent localization of CD31 and GFP in implanted non GFP cultured molars in GFP adult mice showed the origin of neo-formed blood vessels.
    DP: Dental Pulp
    EO: Enamel Organ

    See Abreview

  • This image shows IF using GFP-expressing glial cells (green) transplanted into lesioned rat spinal cord. This was detected using ab6556 anti-GFP antibody and a FITC conjugated secondary antibody. Axons are labelled red by an antibody to neurofilament-200 and a rhodamine secondary. ab6556 reveals the morphology of the transplanted cells to such an extent that their close interactions with axons are obvious. The top picture shows an optical section from a confocal microscope scan showing how a GFP cell wraps around a branched axon travelling longitudinally. The bottom picture consists of an optical section from another confocal scan showing a GFP cell enveloping an axon in the transverse plane. Review by Andrew Toft submitted 19 May 2004.

  • Specific labeling of a Trk-GFP fusion protein being synthesized on ER in sympathetic neurons infected with an adenovirus carrying the construct. The gold is associated with the ER membranes. This was done using a 1/5000 dilution of affinity purified antibody (ab6556). The tissue section was fixed and embedded using durcupan resin.

Anti-GFP antibody (ab6556) 使用論文

This product has been referenced in:
  • Liang D  et al. Therapeutic efficacy of apelin on transplanted mesenchymal stem cells in hindlimb ischemic mice via regulation of autophagy. Sci Rep 6:21914 (2016). ICC/IF . Read more (PubMed: 26902855) »
  • Alvarez-Cienfuegos A  et al. Intramolecular trimerization, a novel strategy for making multispecific antibodies with controlled orientation of the antigen binding domains. Sci Rep 6:28643 (2016). Read more (PubMed: 27345490) »

See all 285 Publications for this product

Product Wall

Based on the sequence homology, I do not think that the jRED antibodies would cross react with GFP and vice versa.

Application Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample Human Tissue sections (Lung carcinoma cells transfected with CMV-GFP, xen)
Antigen retrieval step Heat mediated - Buffer/Enzyme Used: NaCitrate pH 6
Permeabilization No
Specification Lung carcinoma cells transfected with CMV-GFP, xen
Blocking step Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 15% · Temperature: 20°C
Fixative Formaldehyde
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投稿 May 17 2016

Application Immunocytochemistry/ Immunofluorescence
Sample Mouse Cell (Mouse ES cell (YFP) reporter line.)
Permeabilization Yes - 5% donkey serum in PBST (PBS / 0.1% v/v Triton X-100 (Sigma))
Specification Mouse ES cell (YFP) reporter line.
Blocking step 5% donkey serum in PBST (PBS / 0.1% v/v Triton X-100 (Sigma)) as blocking agent for 24 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 4°C
Fixative Paraformaldehyde
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投稿 Sep 21 2015

Application Immunocytochemistry
Sample Human Cell (HEK293T overexpressing GFP)
Permeabilization Yes - 0.1% TRITONX1000.
Specification HEK293T overexpressing GFP
Blocking step BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 1% · Temperature: RT°C
Fixative Formaldehyde
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投稿 May 12 2015

Application Western blot
Loading amount 400000 cells
Gel Running Conditions Reduced Denaturing (12%)
Sample Mouse Cell lysate - whole cell (Mouse Neuroblastoma (Neuro2A))
Specification Mouse Neuroblastoma (Neuro2A)
Treatment 10% FBS and fibroblast growth factor
Blocking step BSA as blocking agent for 2 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 28°C
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投稿 Apr 01 2015

Application Western blot
Loading amount 20 µg
Gel Running Conditions Reduced Denaturing
Sample Human Cell lysate - whole cell (LNCaP cells)
Specification LNCaP cells
Treatment pEGFP-PKD1 transfected
Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 23°C
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投稿 Feb 13 2015

Abreviews
Application Western blot
Loading amount 15 µg
Gel Running Conditions Non-reduced Non-Denaturing (Native) (4-12% Bis-Tris Gel, MES running buffer)
Sample Mouse Tissue lysate - whole (Whole brain tissue lysate)
Specification Whole brain tissue lysate
Blocking step BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
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投稿 Aug 18 2014

Application Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Blocking step Serum as blocking agent for 3 hour(s) and 0 minute(s) · Concentration: 1% · Temperature: RT°C
Antigen retrieval step Heat mediated - Buffer/Enzyme Used: citrate buffer pH6
Sample Mouse Tissue sections (MSC)
Specification MSC
Permeabilization No
Fixative Formaldehyde
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投稿 Apr 24 2014

Application Immunocytochemistry/ Immunofluorescence
Blocking step 2% NDS and 0.1% fish gellatin as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 2% · Temperature: 37°C
Sample Mouse Cell (Medium Spiny Neurons)
Specification Medium Spiny Neurons
Permeabilization No
Fixative Paraformaldehyde
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投稿 Mar 04 2014

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

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投稿 Feb 12 2014

1-10 of 73 Abreviews or Q&A

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