製品の概要

  • 製品名Anti-RFP antibody (Biotin)
    RFP 一次抗体 製品一覧
  • 製品の詳細
    Rabbit polyclonal to RFP (Biotin)
  • 標識Biotin
  • 特異性ab34771 recognises RFP. The antibody is predicted to react with all RFP variants from Discosoma (including DsRed, TdTomato, mOrange etc.) based on sequence homology and polyclonality of the antibody. However, such crossreactivity has not been QC tested.
  • アプリケーション適用あり: ELISA, WB, ICC, IHC-P, IHC-Fr, IHC-FrFlmore details
  • 免疫原

    Red Fluorescent Protein (RFP) fusion protein corresponding to the full length amino acid sequence (234aa) derived from the mushroom polyp coral Discosoma.

  • 特記事項

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

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). Store at -20°C or -80°C. Avoid freeze / thaw cycle.
  • バッファーPreservative: 0.01% Sodium Azide
    Constituents: 10mg/ml BSA, 0.15M Sodium chloride, 0.02M Potassium phosphate. pH 7.2
  • Concentration information loading...
  • 精製度Immunogen affinity purified
  • 特記事項(精製)This product was prepared from monospecific antiserum by immunoaffinity chromatography using Red Fluorescent Protein (Discosoma) coupled to agarose beads followed by solid phase adsorption(s) to remove any unwanted reactivities. No reaction was observed against Human, Mouse or Rat serum proteins. ELISA was used to confirm specificity at less than 0.1% of target signal.
  • 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 ab34771 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
ELISA 1/10000 - 1/80000.
WB 1/2000 - 1/10000. Detects a band of approximately 27 kDa.
ICC 1/1600.
IHC-P 1/1000 - 1/5000. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

Endogenous tissue biotin should be considered as a source of false-positive staining when immunohistochemical or histochemical techniques which use avidin or streptavidin reagents or anti-biotin antibodies as components of the detection system, are applied to tissue sections. We recommned blocking the endogenous biotins.

IHC-Fr 1/400.
IHC-FrFl Use at an assay dependent concentration. PubMed: 24051358

ターゲット情報

  • 関連性Red fluorescent protein cloned from Entacmaea quadricolor eqFP611 (Uniprot IDQ8ISF8)is an important tool in biotechnology and cell biology as a spectrally distinct companion or substitute for the green fluorescent protein (GFP; from Aequorea jellyfish).
  • 別名
    • GFP like chromoprotein antibody
    • red fluorescent protein antibody
    • Red fluorescent protein eqFP611 antibody
    • RFP antibody

Anti-RFP antibody (Biotin) 画像

  • All lanes : Anti-RFP antibody (Biotin) (ab34771) at 1/2500 dilution

    Lane 1 : 10 µg of HeLa cell extract containing GFP recombinant protein
    Lane 2 : 10 µg of a HeLa lysate containing RFP recombinant protein.

    Secondary
    IRDye™800 conjugated Goat anti-Rabbit IgG at 1/5000 dilution

    Observed band size : 27 kDa (why is the actual band size different from the predicted?)
    IRDye™800 fluorescence image was captured using the Odyssey® Infrared Imaging System developed by LI-COR.
  • ab34771 staining mouse subcutaneous tissue that had been injected with a mixture of cells including mouse embryonic stem cells previously transduced with a retrovirus that expresses RFP.

    The sample was fixed with paraformaldehyde and a heat mediated antigen retreival step using Sodium citrate 10 mM (pH 6.0) was performed.  1.5% serum was used as the blocking agent for 30 minutes at 20°C.  The primary antibody was diluted 1/400 in 1xPBS with 1.5% normal serum and incubated for 1 hour at 20°C. A Biotinylated goat anti-rabbit IgG (H+L) antibody was used as the secondary.

    See Abreview

  • ab34771 staining RFP in transfected Mouse N2a cells by ICC (Immunocytochemistry). Cells were fixed with paraformaldehyde, permeabilized with Triton X-100 and blocked with 5% serum for 2 hours at 25°C. Samples were incubated with primary antibody (1/100 in PBS + 2% BSA) for 12 hours at 4°C. An Alexa Fluor®546-Streptavidin -conjugated anti-rabbit IgG polyclonal (1/500) was used as the secondary antibody.

    See Abreview

  • ab34771 staining mouse epidermis that had been damaged and injected with a mixture of cells inlcuding mouse embryonic stem cells previously transduced with a retrovirus that expresses RFP.  The color is developed with DAB.

    The sample was acetone fixed and incubated with ab34771 diluted 1/400 for 1 hour at 20°C. A Biotinylated goat anti-rabbit IgG (H+L) antibody was used as the secondary.

    See Abreview

  • ICC image of ab34771 staining murine embryonic stem cells, transduced with RFP gene, cultured over mouse embryonic fibroblast.  The cells were PFA fixed and blocked with 1.5% serum for 45 minutes at 20°C.  The primary antibody was diluted 1/1600 and incubated for 1 hour and 15 minutes at 20°C. A biotinylated goat anti-rabbit IgG (H+L) was used as the secondary.

    See Abreview

Anti-RFP antibody (Biotin) (ab34771) 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) »
  • Espana-Agusti J  et al. A minimally invasive, lentiviral based method for the rapid and sustained genetic manipulation of renal tubules. Sci Rep 5:11061 (2015). Read more (PubMed: 26046460) »

See all 18 Publications for this product

Product Wall

Thank you for contacting us.

The buffer we have used is Sodium citrate buffer 6.0 pH. The heat mediated antigen retrieval method should be optimized by the end user.

We recommend incubating the sections for 5,10, or 15 minutes in citr...

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The unconjugated antibody is ab124754.
http://www.abcam.com/rfp-antibody-ab124754.html


Thank you for your reply and the detailed information provided. This will be very useful to investigate this case.

It seems that regardless of the blocking conditions used, with the antibody ab62341 you are seeing very high background. This is...

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Thank you for your email.

You are right; I meant the 04/04/13 of course. I can reassure you that the code is valid until 04/04/13.

I wish you good luck with your experiment and look forward to hearing about your experience with this...

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DISCOUNT CODE: ABTRIAL-xxxxx
Expiration date: 04/04/12
Value: €410.00

I am very pleased to hear you would like to accept our offer and test ab34771 in Flow Cytometry. This code will give you €410.00 off your next order before...

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Thank you for your answer.

I would be pleased to offer you the AbTrial for the ab34771 to use this antibody in FlowCytometry. If you decide to order this antibody please let me now in advance so that I can provide you with the code.

Y...

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Thank you for contacting us.

In this case I would recommend to use the Anti-RFP antibody (ab62341). If it is suitable for your experiment you could use a secondary antibody to detect the Anti RFP antibody.

If the conjugate needs to be...

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Thank you for your reply.

We attempt to list all the information that we have about any of our products on the datasheet provided the information is not commercially senstive. Currently we are not aware of ab34771 being tested in transgeni...

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Thank you very much for contacting us with your question.

Ab34771 is conjugated to biotin so that a biotin-streptavidinsystem can be used in the detection of the antibody, which will amplify the signal. To use this antibody in immunofluoresce...

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Thank you for your patience.



I have enquired with the product originator but unfortunately, the actual sequence information isproprietary and cannot be released. They did however mention that the sequence wasderived from thereferen...

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