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  1. Link

    gfp-antibody-ab6556.pdf

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Tags & Cell Markers Fusion / Marker Proteins GFP
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Anti-GFP antibody (ab6556)

  • Datasheet
  • SDS
Reviews (61)Q&A (22)References (1073)

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Immunocytochemistry - Anti-GFP antibody (ab6556)
  • Immunocytochemistry - Anti-GFP antibody (ab6556)
  • Electron Microscopy - Anti-GFP antibody (ab6556)

Key features and details

  • Rabbit polyclonal to GFP
  • Suitable for: IHC-P, Electron Microscopy, ICC, IP, Flow Cyt, IHC-Fr, WB
  • Reacts with: Species independent
  • Isotype: IgG

Get better batch-to-batch reproducibility with a recombinant antibody

Product image
Anti-GFP antibody [EPR14104] (ab183734)
  • Research with confidence – consistent and reproducible results with every batch
  • Long-term and scalable supply – powered by recombinant technology for fast production
  • Success from the first experiment – confirmed specificity through extensive validation
  • Ethical standards compliant – production is animal-free

Overview

  • Product name

    Anti-GFP antibody
    See all GFP primary antibodies
  • Description

    Rabbit polyclonal to GFP
  • Host species

    Rabbit
  • Specificity

    GFP antibody (ab6556) is reactive against all variants of Aequorea victoria GFP such as S65T-GFP, RS-GFP, YFP, CFP, RFP and EGFP.

  • Tested applications

    Suitable for: IHC-P, Electron Microscopy, ICC, IP, Flow Cyt, IHC-Fr, WBmore details
  • Species reactivity

    Reacts with: Species independent
  • Immunogen

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

  • General notes

    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 purified 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.

    Abcam recommended secondaries - Goat Anti-Rabbit HRP (ab205718) and Goat Anti-Rabbit Alexa Fluor® 488 (ab150077).

    See other anti-rabbit secondary antibodies that can be used with this antibody.

    The Life Science industry has been in the grips of a reproducibility crisis for a number of years. Abcam is leading the way in addressing this with our range of recombinant monoclonal antibodies and knockout edited cell lines for gold-standard validation. 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, along with publications, customer reviews and Q&As

Properties

  • Form

    Liquid
  • Storage instructions

    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.
  • Storage buffer

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

    Immunogen affinity purified
  • Purification notes

    This antibody is an affinity purified rabbit anti-GFP antibody purified on an affinity chromatography column made with highly purified recombinant GFP.
  • Clonality

    Polyclonal
  • Isotype

    IgG
  • Research areas

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

Associated products

  • Compatible Secondaries

    • Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077)
  • Isotype control

    • Rabbit IgG, polyclonal - Isotype Control (ChIP Grade) (ab171870)
  • Positive Controls

    • HEK-293 whole cell lysate (ab7902)
  • 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 (ab290)

Applications

The Abpromise guarantee

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.

Application Abreviews Notes
IHC-P (11)
Use at an assay dependent concentration.
Electron Microscopy
1/5000.
ICC (1)
1/2000.
IP (3)
Use at an assay dependent concentration.
Flow Cyt (1)
Use at an assay dependent concentration.
IHC-Fr (4)
Use at an assay dependent concentration.
WB (27)
Use at an assay dependent concentration. Predicted molecular weight: 27 kDa.
Notes
IHC-P
Use at an assay dependent concentration.
Electron Microscopy
1/5000.
ICC
1/2000.
IP
Use at an assay dependent concentration.
Flow Cyt
Use at an assay dependent concentration.
IHC-Fr
Use at an assay dependent concentration.
WB
Use at an assay dependent concentration. Predicted molecular weight: 27 kDa.

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

  • Immunocytochemistry - Anti-GFP antibody (ab6556)
    Immunocytochemistry - Anti-GFP antibody (ab6556)

    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.

  • Immunocytochemistry - Anti-GFP antibody (ab6556)
    Immunocytochemistry - Anti-GFP antibody (ab6556)

    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.

  • Electron Microscopy - Anti-GFP antibody (ab6556)
    Electron Microscopy - Anti-GFP antibody (ab6556)

    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.

Protocols

To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.

Click here to view the general protocols

Datasheets and documents

  • SDS download

  • Datasheet download

    Download

References (1073)

Publishing research using ab6556? Please let us know so that we can cite the reference in this datasheet.

ab6556 has been referenced in 1073 publications.

  • Montenegro-Venegas C  et al. Bassoon inhibits proteasome activity via interaction with PSMB4. Cell Mol Life Sci 78:1545-1563 (2021). PubMed: 32651614
  • Wei H  et al. Clock component OsPRR73 positively regulates rice salt tolerance by modulating OsHKT2;1-mediated sodium homeostasis. EMBO J 40:e105086 (2021). PubMed: 33347628
  • Mukai K  et al. Homeostatic regulation of STING by retrograde membrane traffic to the ER. Nat Commun 12:61 (2021). PubMed: 33397928
  • Phatak V  et al. Mutant p53 promotes RCP-dependent chemoresistance coinciding with increased delivery of P-glycoprotein to the plasma membrane. Cell Death Dis 12:207 (2021). PubMed: 33627632
  • González-Calvo I  et al. Sushi domain-containing protein 4 controls synaptic plasticity and motor learning. Elife 10:N/A (2021). PubMed: 33661101
View all Publications for this product

Customer reviews and Q&As

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

81-83 of 83 Abreviews or Q&A

Question

I will be using the GFP-ab6556(rabbit) antibody on paraffin sections of mouse kidney which will be de-paraffinized prior and the secondary(goat-anti-rabbit) will be peroxidase conjugated. Is this, in your opinion, my best choice in terms of backround and localization?

Read More

Abcam community

Verified customer

Asked on Jan 14 2002

Answer

The system you have chosen should be fine, although you may want to consider using a biotinylated secondary antibody followed by an ABC-peroxidase system. This is a more sensitive system because the biotinylation process is less harsh than conjugation. You should yield a stronger signal with an ABC method, too. Which ever you choose, remember to use in 10% normal goat serum made up in TBS with 1% BSA to block non-specific binding of the secondary. Also use 3% hydrogen peroxide to block endogenous peroxidases. If you choose to use an ABC method, consider incorporating an endogenous biotin block because kidney is a known to be biotin rich.

Read More

Abcam Scientific Support

Answered on Jan 17 2002

Question

I am intrested in using the GFP-ab6556. The question remained to be answered is in some situation which the cells/tissues were not fixed by a fixation, does still the ab recognize eGFP? As far as I know fixation is needed to keep eGFP in tissues after removing the organs from in vivo.

Read More

Abcam community

Verified customer

Asked on Nov 09 2001

Answer

Yes, fixation is necessary to keep eGFP in tissues. However, depending on the nature of your fusion protein, you may not have to fix the tissue. When working with a fusion that is soluble, fixation is advisable.

Read More

Danielle Miller

Abcam Scientific Support

Answered on Nov 14 2001

Question

I have question about your GFP abab6556. Did you change the clone for this Antibody? I ordered this antibody five times since 2006. If you did, when did you change it?

Read More

Abcam community

Verified customer

Asked on Dec 19 2007

Answer

It has been confirmed that this is a rabbit polyclonal antibody directed against highly purified GFP, and is not clone dependent. Ab6556 is purified on a GFP-Sepharose column to yield the final product. Since 2006, antibodies from three different rabbits have been and the most recent batch was from the same rabbit used in sales by Abcam throughout 2007.

Read More

Abcam Scientific Support

Answered on Dec 19 2007

81-83 of 83 Abreviews or Q&A

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