Overview

  • Product name
    GFP ELISA Kit
  • Detection method
    Colorimetric
  • Precision
    Intra-assay
    Sample n Mean SD CV%
    Cell Lysate 9 3.1%
    Inter-assay
    Sample n Mean SD CV%
    Cell Lysate 3 8.2%
  • Sample type
    Cell Lysate, Cell culture media, Tissue Lysate
  • Assay type
    Sandwich (quantitative)
  • Sensitivity
    1.8 pg/ml
  • Range
    2.7 pg/ml - 2000 pg/ml
  • Recovery

    Sample specific recovery
    Sample type Average % Range
    Cell culture media 102 101% - 103%
    Fetal Bovine Serum 97 96% - 98%

  • Assay time
    1h 30m
  • Assay duration
    One step assay
  • Product overview

    Abcam’s GFP in vitro SimpleStep ELISA® (Enzyme-Linked Immunosorbent Assay) kit is designed for the quantitative measurement of GFP protein in cell and tissue lysates.

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

    The SimpleStep ELISA® employs an affinity tag labeled capture antibody and a reporter conjugated detector antibody which immunocapture the sample analyte in solution. This entire complex (capture antibody/analyte/detector antibody) is in turn immobilized via immunoaffinity of an anti-tag antibody coating the well. To perform the assay, samples or standards are added to the wells, followed by the antibody mix. After incubation, the wells are washed to remove unbound material. TMB substrate is added and during incubation is catalyzed by HRP, generating blue coloration. This reaction is then stopped by addition of Stop Solution completing any color change from blue to yellow. Signal is generated proportionally to the amount of bound analyte and the intensity is measured at 450 nm. Optionally, instead of the endpoint reading, development of TMB can be recorded kinetically at 600 nm.

     

  • Notes

    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. YFP differs from GFP due to a mutation at T203Y; antibodies raised against full-length GFP should also detect YFP and other variants.

  • Tested applications
    Suitable for: Sandwich ELISAmore details
  • Platform
    Microplate

Properties

Applications

Our Abpromise guarantee covers the use of ab171581 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
Sandwich ELISA Use at an assay dependent concentration.

Images

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

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

  • Background subtracted data from triplicate measurements are plotted.

Protocols

References

This product has been referenced in:
  • 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). Read more (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). Read more (PubMed: 29349097) »
See all 3 Publications for this product

Customer reviews and Q&As

1-2 of 2 Abreviews or Q&A

Question
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.

Read More

Test for superfold GFP

Excellent Excellent 5/5 (Ease of Use)
Abreviews
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

Submitted Oct 27 2014

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

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