SimpleStep

GFP ELISA Kit (ab171581)

Reviews (1)Q&A (1)References (8)

製品の概要

  • 製品名

    GFP ELISA Kit

  • 検出方法

    Colorimetric

  • 再現性

    Intra-Assay（同時再現性）
    サンプル N 平均値 SD CV%
    Cell Lysate 9 3.1%
    Inter-Assay（日差再現性）
    サンプル N 平均値 SD CV%
    Cell Lysate 3 8.2%

  • サンプルの種類

    Cell Lysate, Cell culture media, Tissue Lysate

  • アッセイタイプ

    Sandwich (quantitative)

  • 検出感度

    1.8 pg/ml

  • 検出範囲

    2.7 pg/ml - 2000 pg/ml

  • 添加回収試験

    特定サンプルでの回収試験
    サンプルの種類 平均 % 測定範囲
    Cell culture media 102 101% - 103%
    Fetal Bovine Serum 97 96% - 98%

  • 全工程の試験時間

    1h 30m

  • ステップ

    One step assay

  • 製品の概要

    GFP ELISA kit (ab171581) is a single-wash 90 min sandwich ELISA designed for the quantitative measurement of GFP protein in cell and tissue lysates. It uses our proprietary SimpleStep ELISA® technology. Quantitate human GFP with 1.8 pg/ml sensitivity.

    SimpleStep ELISA® technology employs capture antibodies conjugated to an affinity tag that is recognized by the monoclonal antibody used to coat our SimpleStep ELISA® plates. This approach to sandwich ELISA allows the formation of the antibody-analyte sandwich complex in a single step, significantly reducing assay time. See the SimpleStep ELISA® protocol summary in the image section for further details. Our SimpleStep ELISA® technology provides several benefits:

            -Single-wash protocol reduces assay time to 90 minutes or less
            -High sensitivity, specificity and reproducibility from superior antibodies
            -Fully validated in biological samples
            -96-wells plate breakable into 12 x 8 wells strips


    Watch our short animated video to see how it works

    A 384-well SimpleStep ELISA® microplate (ab203359) is available to use as an alternative to the 96-well microplate provided with SimpeStep ELISA® kits.


    This kit will detect both the Enhanced and Superfold variants of GFP.



    Accurate ELISAs don't have to take 3 hours or more. SimpleStep ELISA® kits deliver fast results in just 90 minutes by reducing antibody and sample additions to a single step.


  • 特記事項

    Green fluorescent protein (GFP) is a 27 kDa protein derived from the jellyfish Aequorea victoria, which emits green light (emission peak at a wavelength of 509 nm) when excited by blue light (excitation peak at a wavelength of 395 nm). GFP has become an invaluable tool in cell biology research, since its intrinsic fluorescence can be visualized in living cells. GFP fluorescence is stable under fixation conditions and suitable for a variety of applications. GFP has been widely used as a reporter for gene expression, enabling researchers to visualize and localize GFP-tagged proteins within living cells without the need for chemical staining. Other applications of GFP include assessment of protein protein interactions through the yeast two hybrid system and measurement of distance between proteins through fluorescence energy transfer (FRET) protocols. GFP technology has considerably contributed to a greater understanding of cellular physiology. 

    Abcam has not and does not intend to apply for the REACH Authorisation of customers’ uses of products that contain European Authorisation list (Annex XIV) substances.
    It is the responsibility of our customers to check the necessity of application of REACH Authorisation, and any other relevant authorisations, for their intended uses.

  • 試験プラットフォーム

    Microplate

製品の特性

画像

  • Other - GFP ELISA Kit (ab171581)
    Other - GFP ELISA Kit (ab171581)

    SimpleStep ELISA technology allows the formation of the antibody-antigen complex in one single step, reducing assay time to 90 minutes. Add samples or standards and antibody mix to wells all at once, incubate, wash, and add your final substrate. See protocol for a detailed step-by-step guide.

     

  • Example GFP standard curve
    Example GFP standard curve

    Background-subtracted data values (mean +/- SD) are graphed.

  • Example of raw data for GFP standard curve
    Example of raw data for GFP standard curve

    Raw data values are shown in the table

  • GFP cross reactivity data
    GFP cross reactivity data

    This kit recognizes both GFP and enhanced GFP (eGFP) in cell and tissue extracts. No reactivity with mCherry, RFP or YFP was observed.

  • Titration of GFP spiked 143B cell lysate within the working range of the assay
    Titration of GFP spiked 143B cell lysate within the working range of the assay

    Background subtracted data from triplicate measurements are plotted.

  • AbReview
    AbReviewThis image is courtesy submitted by Ling Chun

    The kit works with superfold GFP. 

    See Abreview

  • Linearity of dilution for spiked GFP
    Linearity of dilution for spiked GFP

    GFP spiked 143B lysate was measured in 3-fold dilution series

参考文献

ab171581 を使用した論文を発表された方は、こちらまでお知らせください。データシートに掲載させていただきます。

ab171581 は 8 報の論文で使用されています。

  • Anjo SI  et al. A different vision of translational research in biomarker discovery: a pilot study on circulatory mitochondrial proteins as Parkinson's disease potential biomarkers. Transl Neurodegener 9:11 (2020). PubMed: 32266064
  • Lin CJ  et al. Microbubble-facilitated ultrasound pulsation promotes direct a-synuclein gene delivery. Biochem Biophys Res Commun 517:77-83 (2019). PubMed: 31327496
  • Yu K  et al. Proof of principle: Physiological transfer of small numbers of bacteria from mother to fetus in late-gestation pregnant sheep. PLoS One 14:e0217211 (2019). PubMed: 31170184
  • Ding K  et al. AAV8-vectored suprachoroidal gene transfer produces widespread ocular transgene expression. J Clin Invest 129:4901-4911 (2019). PubMed: 31408444
  • Hori A  et al. Unexpected role of the IMD pathway in Drosophila gut defense against Staphylococcus aureus. Biochem Biophys Res Commun 495:395-400 (2018). PubMed: 29108998
  • Nass SA  et al. Universal Method for the Purification of Recombinant AAV Vectors of Differing Serotypes. Mol Ther Methods Clin Dev 9:33-46 (2018). PubMed: 29349097
  • Reid CA  et al. Development of an inducible anti-VEGF rAAV gene therapy strategy for the treatment of wet AMD. Sci Rep 8:11763 (2018). ELISA . PubMed: 30082848
  • Wandrey G  et al. Probing unnatural amino acid integration into enhanced green fluorescent protein by genetic code expansion with a high-throughput screening platform. J Biol Eng 10:11 (2016). PubMed: 27733867

レビューと Q&A

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1-2 of 2 Abreviews or Q&A

Question

Will this work on GFP fusion proteins?

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Answer

We have not tested ab171581 with a GFP fusion protein (purified or in cell extract) before. The epitopes for the capture and detector antibodies are also not known because both are made using full length immunogens. However, both antibodies alone have detected GFP fusion proteins in other applications (WB, IP), so it is encouraging that at least the individual antibodies alone are capable of detecting GFP fusion proteins.

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Test for superfold GFP

 Excellent Excellent 5/5 (Ease of Use)
Abreviews
abreview image
The kit works with superfold GFP. I have tested again our purified superfold GFP protein and also some cell lysate from cells expressing superfold GFP. Both worked.

When compared with the GFP stock provided by the kit, the sensitivity against superfold GFP is lower than GFP. This could be due to our protein stock is stored at -80C for sometime.

Ling Chun

Verified customer

投稿 Oct 27 2014

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