製品の概要

  • 製品名Anti-GFP antibody (Biotin)
    GFP 一次抗体 製品一覧
  • 製品の詳細
    Goat polyclonal to GFP (Biotin)
  • 標識Biotin
  • 特異性Antibody recognizes wild type, recombinant and enhanced forms of GFP (EGFP). No reaction was observed against Human, Mouse and Rat Serum Proteins.
  • アプリケーション適用あり: WB, IP, ICC/IF, Sandwich ELISA, IHC-Pmore details
  • 免疫原

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

    MSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLKFICTT GKLPVPWPTL VTTFSYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTI FFKDDGNYKTRAEVKFEGDTLV NRIELKGIDFKEDGNILGHKLEYNYN SHNVYIMADKQKNGIKVNFKIRHNIEDGSVQLAD HYQQNTPIGDGPVL LPDNHYLSTQSALSKDPNEKRDHMVLLEFVTAAGITHGMDELYK

  • 特記事項

    Designed to detect GFP and its variants in ELISA (sandwich or capture), immunoblotting and immunoprecipitation.

    Biotinamidocaproate N-Hydroxysuccinimide Ester (BAC) Biotin/Protein Ratio: 10-20 BAC molecules per goat IgG molecule.

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. Avoid freeze / thaw cycle.
  • バッファーpH: 7.20
    Preservative: 0.01% Sodium azide
    Constituents: 0.44% Potassium phosphate, 0.88% Sodium chloride

    10 mg/mL BSA, immunoglobulin and protease free
  • Concentration information loading...
  • 精製度IgG fraction
  • 特記事項(精製)This product 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.
  • 一次抗体 備考Designed to detect GFP and its variants in ELISA (sandwich or capture), immunoblotting and immunoprecipitation.
  • 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 ab6658 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
WB 1/2000 - 1/10000.
IP Use at an assay dependent concentration.
ICC/IF 1/1000 - 1/5000.
Sandwich ELISA 1/10000 - 1/50000. Can be paired for Sandwich ELISA with Mouse monoclonal [9F9.F9] to GFP (ab1218).

May be used as capture or detection antibody in a Sandwich ELISA.

IHC-P 1/250.

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 (Biotin) 画像

  • Anti-GFP antibody (Biotin) (ab6658)

    Additional bands at : 28 kDa. We are unsure as to the identity of these extra bands.
  • ab6658 staining GFP in human melanoma cells recovered from nude mice by Immunocytochemistry/ immunoflurescence. Cells were fixed with formaldehyde, permeabilized with 0.25% Triton X-100 RT for 10min and blocking with commercially available blocking buffer was performed for 30 minutes at RT. Samples were incubated with primary antibody (1:50) for 18 hours at 4°C. An Alexa Fluor®488-conjugated donkey polyclonal to goat IgG was used as secondary antibody at 1/100 dilution. Green color indicates GFP/Fibrolast cells, while red color indicates Ki67 positive cells, most of them are tumor cells (Abcam`s ab15580 was used for the detection).

    See Abreview

  • Immunofluorescence Microscopy using ab6658. 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 min 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.

  • Immunofluorescence Microscopy using ab6658. 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. Primary antibody: 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 min 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.

  • ab6658 staining GFP in rat brain cells infected with viruses containing GFP under a CMV promoter by Immunocytochemistry/ Immunofluorescence.Cells were fixed with formaldehyde, permeabilized using 0.2% Triton, blocked with 20% serum and then incubated with ab6658 at a 1:50 dilution for 20 hours at 25°C. The secondary used was an Alexa-Fluor 488 conjugated rabbit polyclonal, used at a 1/200 dilution.

    See Abreview

Anti-GFP antibody (Biotin) (ab6658) 使用論文

This product has been referenced in:
  • Bando JK  et al. Identification and distribution of developing innate lymphoid cells in the fetal mouse intestine. Nat Immunol 16:153-60 (2015). IHC-Fr . Read more (PubMed: 25501629) »
  • Volpetti F  et al. A microfluidic platform for high-throughput multiplexed protein quantitation. PLoS One 10:e0117744 (2015). Read more (PubMed: 25680117) »

See all 26 Publications for this product

Product Wall



I am sorry to confirm that the binding constant is not often determined for research grade antibodies and there is no binding constant information available for this particular product.

I am sorry we have no data to share to help ans...

Read More


I performed the following molecular weight calculations for this antibody:

Assuming 15 biotin molecules per IgG molecule:

MW IgG = 150 kDa
MW biotin = 244.31 g/mol = 244.31 Da = 0.24431 kDa
MW ab6658 = 150 + (15*0.2...

Read More

We do not determine the percentage of molecules conjugated to biotin, but it is expected to be close to 100%. The conjugation ratio between our biotin and IgG is usually 5-6 molecules of biotin per molecule of IgG.

The unconjugated version of ab6658 is ab6673.

http://www.abcam.com/GFP-antibody-ab6673.html

Thank you very much for contacting us with your question.

In general, using a secondary antibody will help amplify the signal from the primary antibody. With a pre-conjugated primary antibody, the signal may be weaker than using a conjugated ...

Read More

>pecies specificity is not applicable for this product. You can try this antibody for detection of GFP in any type of cells if they express GFP protein.

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Immunocytochemistry/ Immunofluorescence
Sample Rat Cell (Brain tissue slices)
Specification Brain tissue slices
Fixative Formaldehyde
Permeabilization Yes - Triton 0.2%
Blocking step Serum as blocking agent for 1 hour(s) and 30 minute(s) · Concentration: 20% · Temperature: 25°C
Username

Dr. Efrat Shema

Verified customer

投稿 Aug 04 2011

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample Rat Tissue sections (Brain)
Specification Brain
Fixative Paraformaldehyde
Antigen retrieval step Heat mediated - Buffer/Enzyme Used: Sodium citrate pH 6.0
Permeabilization Yes - Triton 0.2%
Blocking step Serum as blocking agent for 1 hour(s) and 30 minute(s) · Concentration: 20%
Username

Dr. Efrat Shema

Verified customer

投稿 Aug 02 2011

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample Human Tissue sections (Human melanoma tumor recovered from nude mice.)
Specification Human melanoma tumor recovered from nude mice.
Fixative Formaldehyde
Antigen retrieval step Heat mediated - Buffer/Enzyme Used: 10mm Tris, 1mM EDTA, 0.5%Tween20, pH9.0, 95-100C 40min
Permeabilization Yes - 0.25% Triton X-100 RT 10min
Blocking step Dako Cytomation Serum Free General Blocking Cat#X0909 as blocking agent for 30 minute(s) · Concentration: 0% · Temperature: RT°C
Username

Hongwei Shao

Verified customer

投稿 Apr 19 2010

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Western blot
Sample Mouse Tissue lysate - whole (lung)
Loading amount 25 µg
Specification lung
Gel Running Conditions Reduced Denaturing
Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 37°C
Username

Abcam user community

Verified customer

投稿 Mar 10 2009

1-10 of 16 Abreviews or Q&A

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