Overview

  • Product name
    Anti-GFP antibody [9F9.F9]
    See all GFP primary antibodies
  • Description
    Mouse monoclonal [9F9.F9] to GFP
  • Host species
    Mouse
  • Specificity
    Assayed by ELISA for direct binding of antigen recognizes wild type and recombinant GFP. Well suited to titrate GFP in solution using either form of the antibody as the capture or detection antibodies. Shows no reactivity against red fluorescence protein (RFP). This antibody is known to cross react with the wild type (wt), recombinant (rGFP) and enhanced (eGFP) forms.
  • Tested applications
    Suitable for: WB, IHC-Fr, Sandwich ELISA, ICC/IF, IPmore details
  • Immunogen

    Fusion protein corresponding to GFP aa 1-246. Full Length Fusion Protein. Derived from the jellyfish Aequorea victoria.
    Database link: P42212

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: 7.20
    Preservative: 0.01% Sodium azide
    Constituents: 0.87% Sodium chloride, 0.42% Potassium phosphate
  • Concentration information loading...
  • Purity
    Protein A purified
  • Purification notes
    GFP Monoclonal Antibody was prepared from tissue culture supernatant by Protein A affinity chromatography.
  • Clonality
    Monoclonal
  • Clone number
    9F9.F9
  • Isotype
    IgG1
  • Light chain type
    kappa
  • Research areas

Applications

Our Abpromise guarantee covers the use of ab1218 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
WB 1/3000 - 1/30000.

(See Abreviews). Recognises the immunogen under native conditions and reduced/denatured conditions.

IHC-Fr 1/1000 - 1/5000.

Works also with eYFP.

Sandwich ELISA 1/80000 - 1/500000. Can be paired for Sandwich ELISA with Goat polyclonal to GFP (Biotin) (ab6658).

ab1218 can be used to detect GFP by ELISA (sandwich or capture) for the direct binding of antigen. Biotin conjugated monoclonal anti-GFP is well suited to titrate GFP in a sandwich ELISA in combination with polyclonal anti-GFP as a capture antibody.

ICC/IF Use at an assay dependent concentration.
IP Use at an assay dependent concentration. PubMed: 16135826

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

  • ab1218 staining GFP in Monkey Kidney cells by ICC/IF (Immunocytochemistry/immunofluorescence). Cells were fixed with formaldehyde, permeabilized with 1% Triton X-100 and blocked with 3% BSA for 1 hour at 25°C. Samples were incubated with primary antibody (1/500) for 12 hour at 4°C. An undiluted Alexa Fluor® 488-conjugated Goat polyclonal was used as the secondary antibody

    See Abreview

  • ab1218 staining GFP in human prostate epithelial primary cells by Immunocytochemistry/ Immunofluorescence. Cells (ARPE-19 cells transfected with mouse Rab27A-GFP) were fixed with paraformaldehyde, permeabilized with 0.5% Triton x100 and blocking with 5% serum was performed for 20 minutes at 25°C. Samples were incubated with primary antibody (1/500: in 1% goat serum, 0.1%TX100, 1XPBS) for 16 hours at 4°C. An Alexa Fluor®546-conjugated goat polyclonal to mouse IgG was used as secondary antibody at 1/500 dilution. DAPI was used to stain the cell nuclei (blue) and mRab27A-GFP with ab1218 (red).

    See Abreview

  • All lanes : Anti-GFP antibody [9F9.F9] (ab1218) at 1 mg/ml (1 hour at room temperature)

    Lane 1 : HeLa lysate 50ng with BLOTTO, overnight at 4°C
    Lane 2 : HeLa lysate 100ng with BLOTTO, overnight at 4°C
    Lane 3 : HeLa lysate 500ng with BLOTTO, overnight at 4°C

    Blocking peptides at 5 % per lane.

    Secondary
    All lanes : IRDye® 800 conjugated Goat anti-mouse IgG (H+L), 45 minutes at room temperature at 1/2500 dilution

    Additional bands at: 27 kDa. We are unsure as to the identity of these extra bands.

  • All lanes : Anti-GFP antibody [9F9.F9] (ab1218) at 1/1000 dilution (2 hours at 25°C)

    Lane 1 : J774 with Milk, 2 hours at 25°C
    Lane 2 : J774+pEGFP-N1 with Milk, 2 hours at 25°C

    Lysates/proteins at 20 µg per lane.

    Blocking peptides at 5 % per lane.

    Secondary
    All lanes : Goat anti-mouse HRP at 1/10000 dilution

    Developed using the ECL technique.

    Performed under reducing conditions.

    Additional bands at: 26 kDa. We are unsure as to the identity of these extra bands.


    Exposure time: 1 minute

    See Abreview

  • Immunoprecipitation using ab1218 at 0.01µl/µl HEK (human embryonic kidney) 293 whole cell lysate incubated with protein G matrix for 1 hour at 4°C.

    Total protein input was 3,000,000 cells.

    Lane 1: GFP-RhoA wt

    Lane 2: GFP-Rac1 wt

    Lane 3: GFP-empty

    The western blot antibody was ab290 (undiluted).

    See Abreview

References

This product has been referenced in:
  • Wu N  et al. RCC2 over-expression in tumor cells alters apoptosis and drug sensitivity by regulating Rac1 activation. BMC Cancer 18:67 (2018). Read more (PubMed: 29321004) »
  • Mazzon M  et al. Alphavirus-induced hyperactivation of PI3K/AKT directs pro-viral metabolic changes. PLoS Pathog 14:e1006835 (2018). Read more (PubMed: 29377936) »

See all 180 Publications for this product

Customer reviews and Q&As

We do not have data for reactivity of ab1218 with acGFP but a customer sent a positive review of ab290 for IP and WB of acGFP and acGFP-tagged p27. Here is the URL of the review http://www.abcam.com/GFP-antibody-ChIP-Grade-ab290/reviews/33378 .

Thank you for contacting us. Yes, this antibody does recognize eGFP.

This mouse monoclonal antibody against GFP was raised against a full length recombinant protein. Unfortunately, the specific epitope has not been mapped, therefore we do not know the exact sequence recognized by the antibody.

We have not tested ab1218 for reactivity with BFP. It is possible that this antibody may react with BFP, but as we have not mapped the epitope for this antibody, I do not have any further information about whether this antibody would be likely to react...

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Thank you very much for your interest in ab1218.

With this testing discount program, you do not return the antibody for a refund if it doesn't work. Instead, you would earn a discount code good for a free antibody off a future purchase (buy-o...

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This monoclonal hasn't been tested against eYFP or mCherry, however it is expected to bind to eYFP but not to mCherry since the sequence of mCherry is quite different from YFP. But again, this hasn't been tested.


If you would like to te...

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Todos nuestros productos son estables durante alrededor de un año si se conservan a 4C y hasta cinco años conservados a -20C. Si se almacenan a -80 pueden durar hasta 10 años.

Los anticuerpos mencionados no han podido sufrir daño alguno por e...

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>I am sorry to confirm that these GFP antibodies have not been tested for detection of GFP from Zoanthus sp.

Our GFP antibodies, including ab6556, ab1218 and ab290are all raised against immunogens from the Aquorea victoria GFP protein, which...

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I can confirm that ab1218 can also be used for detection of YFP protein .

Thank you for confirming these details and for your cooperation. The details provided enable us to closely monitor the quality of our products. I am sorry this product did not perform as stated on the datasheet and for the inconvenience this has ca...

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1-10 of 26 Q&A

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

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