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    fitc-gfp-antibody-ab6662.pdf

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Tags & Cell Markers Fusion / Marker Proteins GFP
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FITC Anti-GFP antibody (ab6662)

  • Datasheet
Reviews (14)Q&A (7)References (125)

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Western blot - FITC Anti-GFP antibody (ab6662)
  • Immunofluorescence - FITC Anti-GFP antibody (ab6662)
  • Immunofluorescence - FITC Anti-GFP antibody (ab6662)
  • Immunohistochemistry (Frozen sections) - FITC Anti-GFP antibody (ab6662)
  • Immunohistochemistry (Frozen sections) - FITC Anti-GFP antibody (ab6662)
  • Immunohistochemistry (Frozen sections) - FITC Anti-GFP antibody (ab6662)

Key features and details

  • FITC Goat polyclonal to GFP
  • Suitable for: IHC-FoFr, IHC-Fr, WB, ICC/IF
  • Reacts with: Species independent
  • Conjugation: FITC. Ex: 493nm, Em: 528nm
  • Isotype: IgG

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Overview

  • Product name

    FITC Anti-GFP antibody
    See all GFP primary antibodies
  • Description

    FITC Goat polyclonal to GFP
  • Host species

    Goat
  • Conjugation

    FITC. Ex: 493nm, Em: 528nm
  • Tested applications

    Suitable for: IHC-FoFr, IHC-Fr, WB, ICC/IFmore details
  • Species reactivity

    Reacts with: Species independent
  • Immunogen

    Recombinant full length protein corresponding to GFP aa 1-246.
    Sequence:

    MSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLKFICTT GKLPVPWPTLVTTFSYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFF KDDGNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNV YIMADKQKNGIKVNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHY LSTQSALSKDPNEKRDHMVLLEFVTAAGITHGMDELYK


    Database link: P42212
    Run BLAST with BLAST the sequence with ExPASy Run BLAST with BLAST the sequence with NCBI
  • Positive control

    • WB: Recombinant A. victoria GFP protein.
  • General notes

    Designed to detect GFP and its variants in ELISA (sandwich or capture), immunoblotting and immunoprecipitation. Fluorescein conjugated anti-GFP was assayed by immunofluorescence microscopy on prokaryotic (E.coli) and eukaryotic (CHO cells) expression systems and was shown to detect GFP containing inserts. Significant amplification of signal was detected using fluorochrome conjugated anti-GFP relative to the fluorescence of GFP alone.
    In case of unexpected background, use pre-adsorbed secondary antibodies.

    Fluorescein isothiocyanate (FITC) (MW 390 daltons) Absorption Wavelength: 495 nm Emission Wavelength: 528 nm Fluorochrome/Protein Ratio: 3.5 moles FITC per mole of Goat IgG

    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.

Properties

  • Form

    Liquid
  • Storage instructions

    Shipped at 4°C. Store at +4°C short term (1-2 weeks). Store at -20°C. Avoid freeze / thaw cycle.
  • Storage buffer

    Preservative: 0.01% Sodium azide
    Constituents: 0.42% Potassium phosphate, 0.87% Sodium chloride, 1% BSA

    BSA Immunoglobulin and Protease free
  • Concentration information loading...
  • Purity

    Protein G purified
  • Purification notes

    GFP Fluorescein Conjugated Antibody was prepared from monospecific antiserum by immunoaffinity chromatography using Green Fluorescent Protein (Aequorea victoria) coupled to agarose beads followed by solid phase adsorption(s) to remove any unwanted reactivities.
  • Primary antibody notes

    Designed to detect GFP and its variants in ELISA (sandwich or capture), immunoblotting and immunoprecipitation. Fluorescein conjugated anti-GFP was assayed by immunofluorescence microscopy on prokaryotic (E.coli) and eukaryotic (CHO cells) expression systems and was shown to detect GFP containing inserts. Significant amplification of signal was detected using fluorochrome conjugated anti-GFP relative to the fluorescence of GFP alone.
  • Clonality

    Polyclonal
  • Isotype

    IgG
  • Research areas

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

Associated products

  • Isotype control

    • FITC Goat IgG - Isotype control (ab37374)
  • 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)

Applications

Our Abpromise guarantee covers the use of ab6662 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-FoFr 1/250.
IHC-Fr Use at an assay dependent concentration.
WB 1/10000 - 1/100000.
ICC/IF 1/200 - 1/400.
IF 1/500 - 1/2500.

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

  • Western blot - FITC Anti-GFP antibody (ab6662)
    Western blot - FITC Anti-GFP antibody (ab6662)
    FITC Anti-GFP antibody (ab6662) + Recombinant A. victoria GFP protein (ab84191)

    Secondary
    Fluorescein goat secondary antibody for 60 minutes at RT at 1/1000 dilution


    Block: Blocking buffer for 30 minutes at RT.

  • Immunofluorescence - FITC Anti-GFP antibody (ab6662)
    Immunofluorescence - FITC Anti-GFP antibody (ab6662)Kim at el PLoS Genet. 2018 Feb 8;14(2):e1007204. doi: 10.1371/journal.pgen.1007204. eCollection 2018 Feb. Fig 4. Reproduced under the Creative Commons license http://creativecommons.org/licenses/by/4.0/

    Oscillation of Cyclin E and E2F target gene expression is deregulated in de2f1b mutant salivary glands.

    Salivary glands of control and de2f1b mutant early (80–85 hr AEL) third instar larvae expressing PCNA-GFP (green, ab6662) are stained with anti-dE2F1 (red). The region where high PCNA-GFP is observed with low dE2F1 is marked by an asterisk.

    For Immunostaining, third instar imaginal discs and salivary glands were dissected in PBS and immediately fixed in 4% formaldehyde in PBS for 20 minutes at room temperature with the exception of tissues subjected to anti-dE2F1 staining that were fixed for 30 minutes on ice. Fixed tissues were then washed with 0.3% PBST (0.3% TritonX-100 in 1XPBS) and 0.1% PBST (0.1% TritonX-100 in 1XPBS). Samples were incubated with appropriate amount of primary antibody in 0.1% PBST and 1%BSA overnight. Samples were then washed with 0.1% PBST, incubated in secondary antibody in 0.1% PBST and 1% BSA for 2 hours, followed by several washes in 0.1% PBST prior to mounting.

  • Immunofluorescence - FITC Anti-GFP antibody (ab6662)
    Immunofluorescence - FITC Anti-GFP antibody (ab6662)

    Immunofluorescence Microscopy using ab6662.

    Tissue: Drosophila melanogaster late stage embryonic central nervous system.

    Fixation: 0.5% PFA.

    Antigen retrieval: Not required.

    Primary antibody: Anti-GFP antibody at a 1/1,000 for 1 h at RT.

    Secondary antibody: AlexaFluor 488™ conjugated anti-Goat antibody at 1/300 for 45 minutes at RT.

    Panel A: shows a lateral view (ventral left).

    Panels B and C: shows ventral views of whole mount embryos at 63x magnification (plus 2x digital zoom).

    In all panels, anterior is up.

    Staining: tau-GFP cell bodies (large arrowhead) and axons of motorneurons (arrow) and interneurons (small arrowhead) as green fluorescent signal.

  • Immunohistochemistry (Frozen sections) - FITC Anti-GFP antibody (ab6662)
    Immunohistochemistry (Frozen sections) - FITC Anti-GFP antibody (ab6662)This image is courtesy of an Abreview submitted by Dr Joshua Breunig

    ab6662 staining mouse brain tissue sections (inducible GFP reporter) by IHC-Fr.

    The tissue was paraformaldehyde fixed and blocked with serum and then incubated with the antibody at a 1/1000 dilution for 1 hour.

    Staining is shown in the left hand panel. The middle panel shows staining with a rabbit anti-GFP antibody and the right hand panel shows the merged images (plus DAPI).

    ab6662 gives no noticable background and it is found that when viewing on an epifluorescent the exposure time is significantly reduced.

    See Abreview

  • Immunohistochemistry (Frozen sections) - FITC Anti-GFP antibody (ab6662)
    Immunohistochemistry (Frozen sections) - FITC Anti-GFP antibody (ab6662)

    Immunofluorescence Microscopy using ab6662.

    Tissue: Sf-1:Cre mice crossed to the Z/EG reporter line. Mouse brain (coronal view, 20X magnification).

    Fixation: 4% PFA/PBS with o/n fixation, and subsequently transferred to a 30% sucrose solution.

    Antigen retrieval: Frozen in OCT freezing medium (Sakura) and cryostat sectioned at 40 microns.

    Goat anti-GFP was used at 1/500 dilution in free floating imunnohistochemistry to detect GFP.

    Secondary antibody: Fluorchrome conjugated Anti-goat IgG secondary antibody was used for detection at 1:500 at 1/10,000 for 45 minutes at RT.

    Localization: Sf-1+ neurons and their processes of the ventromedial nucleus of the hypothalamus.

    Staining: eGFP as green fluorescent signal and sections were counterstained with DAPI.

  • Immunohistochemistry (Frozen sections) - FITC Anti-GFP antibody (ab6662)
    Immunohistochemistry (Frozen sections) - FITC Anti-GFP antibody (ab6662)

    These pictures show confocal immunofluorescence using GFP-expressing glial cells (green) transplanted into the lesioned rat spinal cord.

    Detected using ab6662 and a standard FITC filter set. Axons are labeled red by an antibody to neurofilament-200 and a rhodamine secondary antibody. The upper panel shows the centre of the transplant site at low power. Numerous GFP-positive cells can be seen mingling with axons. The lower panel shows, at high power in a single optical section, how ab6662 reveals the morphology of the transplanted cells to such an extent that their close interactions with axons are obvious - the cell depicted can be seen wrapping around a neurofilament-200 positive axon.

    These images were kindly supplied as part of the review submitted by Andrew Toft.

Protocols

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

Click here to view the general protocols

Datasheets and documents

    • Datasheet
  • References (125)

    Publishing research using ab6662? Please let us know so that we can cite the reference in this datasheet.

    ab6662 has been referenced in 125 publications.

    • Maurer GW  et al. Analysis of genes within the schizophrenia-linked 22q11.2 deletion identifies interaction of night owl/LZTR1 and NF1 in GABAergic sleep control. PLoS Genet 16:e1008727 (2020). PubMed: 32339168
    • Gao L  et al. Vps35 Deficiency Impairs Cdk5/p35 Degradation and Promotes the Hyperphosphorylation of Tau Protein in Retinal Ganglion Cells. Invest Ophthalmol Vis Sci 61:1 (2020). PubMed: 31995153
    • Smith SJ  et al. Evolutionary expansion of apical extracellular matrix is required for the elongation of cells in a novel structure. Elife 9:N/A (2020). PubMed: 32338602
    • Vitre B  et al. IFT proteins interact with HSET to promote supernumerary centrosome clustering in mitosis. EMBO Rep 21:e49234 (2020). PubMed: 32270908
    • Comella-Bolla A  et al. Human Pluripotent Stem Cell-Derived Neurons Are Functionally Mature In Vitro and Integrate into the Mouse Striatum Following Transplantation. Mol Neurobiol 57:2766-2798 (2020). PubMed: 32356172
    View all Publications for this product

    Customer reviews and Q&As

    Show All Reviews Q&A
    Submit a review Submit a question

    1-10 of 21 Abreviews or Q&A

    Immunohistochemistry (Frozen sections) abreview for Anti-GFP antibody (FITC)

    Excellent
    Abreviews
    Abreviews
    abreview image
    Application
    Immunohistochemistry (Frozen sections)
    Sample
    Mouse Tissue sections (bone marrow)
    Permeabilization
    No
    Specification
    bone marrow
    Blocking step
    Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 10% · Temperature: RT°C
    Fixative
    Paraformaldehyde
    Read More

    Abcam user community

    Verified customer

    Submitted Sep 16 2019

    Immunocytochemistry/ Immunofluorescence abreview for Anti-GFP antibody (FITC)

    Good
    Abreviews
    Abreviews
    abreview image
    Application
    Immunocytochemistry/ Immunofluorescence
    Sample
    Mouse Cell (Epithelial cells)
    Permeabilization
    Yes - Triton 100
    Specification
    Epithelial cells
    Blocking step
    BSA as blocking agent for 30 minute(s) · Concentration: 5% · Temperature: 25°C
    Fixative
    Paraformaldehyde
    Read More

    Dr. Tina Yuan

    Verified customer

    Submitted Sep 16 2019

    Immunohistochemistry (Frozen sections) abreview for Anti-GFP antibody (FITC)

    Excellent
    Abreviews
    Abreviews
    abreview image
    Application
    Immunohistochemistry (Frozen sections)
    Sample
    Mouse Tissue sections (Bone)
    Permeabilization
    No
    Specification
    Bone
    Blocking step
    Vetor Blocker as blocking agent for 30 minute(s) · Concentration: 1% · Temperature: 23°C
    Fixative
    Paraformaldehyde
    Read More

    Abcam user community

    Verified customer

    Submitted Sep 16 2019

    Western blot abreview for Anti-GFP antibody (FITC)

    Average
    Abreviews
    Abreviews
    Application
    Western blot
    Sample
    Mouse Tissue lysate - nuclear (Bone)
    Gel Running Conditions
    Reduced Denaturing
    Loading amount
    20 µg
    Specification
    Bone
    Blocking step
    BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
    Read More

    Dr. Tina Yuan

    Verified customer

    Submitted Aug 28 2019

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

    Average
    Abreviews
    Abreviews
    abreview image
    Application
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
    Sample
    Mouse Tissue sections (Long bone)
    Antigen retrieval step
    Heat mediated - Buffer/Enzyme Used: Antigen Unmasking Solution, Citric Acid Based
    Permeabilization
    Yes - Triton 100
    Specification
    Long bone
    Fixative
    Paraformaldehyde
    Read More

    Dr. Tina Yuan

    Verified customer

    Submitted Aug 28 2019

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

    Excellent
    Abreviews
    Abreviews
    abreview image
    Application
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
    Sample
    Mouse Tissue sections (Retina)
    Antigen retrieval step
    Heat mediated - Buffer/Enzyme Used: citrate buffer
    Permeabilization
    No
    Specification
    Retina
    Blocking step
    Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 10% · Temperature: 25°C
    Fixative
    Davidson
    Read More

    Abcam user community

    Verified customer

    Submitted Apr 08 2019

    Immunohistochemistry free floating abreview for Anti-GFP antibody (FITC)

    Excellent
    Abreviews
    Abreviews
    abreview image
    Application
    Immunohistochemistry free floating
    Sample
    Mouse Tissue sections (brain)
    Specification
    brain
    Read More

    James Bogenpohl

    Verified customer

    Submitted Jul 16 2018

    Immunohistochemistry (Frozen sections) abreview for Anti-GFP antibody (FITC)

    Excellent
    Abreviews
    Abreviews
    abreview image
    Application
    Immunohistochemistry (Frozen sections)
    Sample
    Mouse Tissue sections (Intestine)
    Permeabilization
    No
    Specification
    Intestine
    Blocking step
    BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5%
    Fixative
    Paraformaldehyde
    Read More

    Abcam user community

    Verified customer

    Submitted Dec 18 2012

    Question


    je travaille sur une nouvelle application en cytométrie de flux en
    neurosciences. J'aurais besoin d'un anticorps anti-GFP (fait dans la
    chèvre ou l'âne, mais pas dans le lapin, la souris ou le poulet) déjà
    couplé à du FITC. J'ai vu que vous disposiez d'une référence (ab6662)
    qui correspond à ces critères mais elle n'est pas validée en
    cytométrie de flux, et je ne peux pas me permettre de commander un
    anticorps sans aucune certitude sur son bon fonctionnement. Je ne
    connais pas vos politiques commerciales, mais serait-il envisageable
    de tester un petit aliquot (10µL devraient suffire pour valider
    l'anticorps), puis si cela fonctionne, vous envoyer les histogrammes
    de cytométrie et effectuer une commande pour réaliser mes expériences?
    Bien cordialement,

    Read More

    Abcam community

    Verified customer

    Asked on Dec 12 2012

    Answer

    Ab6662 n’a effectivement pas encore été testé en cytométrie en flux et nous n’offrons pas d’échantillon gratuit à des fins de tests préliminaires. Cependant, si vous souhaitez tester cet anticorps en cytométrie en flux, il me sera possible de vous offrir une remise. Après soumission d’une Abreview avec vos résultats, un avoir de la valeur de l'anticorps testé vous sera offert.

    Étapes à suivre :

    1. Nous informer que vous souhaitez tester l’anticorps en cytométrie en flux et nous vous enverrons un code promotionnel (valable 4 mois) à utiliser après obtention et envoi de vos résultats,

    2. Acheter l'anticorps,

    3. Nous envoyer vos résultats (positifs ou négatifs) obtenus sous forme d'Abreview (www.abcam.com/abreviews).

    4. L'envoi de vos résultats activera le code et vous donnera un avoir de la valeur de l'anticorps testé.

    J'espère que ces informations vous aideront. N'hésitez pas à me contacter si vous avez besoin de plus de renseignements.

    Read More

    Abcam Scientific Support

    Answered on Dec 12 2012

    Immunocytochemistry/ Immunofluorescence abreview for Anti-GFP antibody (FITC)

    Good
    Abreviews
    Abreviews
    abreview image
    Application
    Immunocytochemistry/ Immunofluorescence
    Sample
    Mouse Cell (Intestine)
    Specification
    Intestine
    Fixative
    Paraformaldehyde
    Permeabilization
    No
    Blocking step
    BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 23°C
    Read More

    Abcam user community

    Verified customer

    Submitted Apr 17 2012

    1-10 of 21 Abreviews or Q&A

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