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  1. Link

    products/elisa/gfp-elisa-kit-ab171581.pdf

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Tags & Cell Markers Fusion / Marker Proteins GFP
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SimpleStep

GFP ELISA Kit (ab171581)

  • Datasheet
  • SDS
  • Protocol Booklet
Reviews (3)Q&A (1)References (14)

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Other - GFP ELISA Kit (ab171581)
  • Example GFP standard curve
  • Example of raw data for GFP standard curve
  • GFP cross reactivity data
  • Titration of GFP spiked 143B cell lysate within the working range of the assay
  • AbReview
  • Linearity of dilution for spiked GFP

Key features and details

  • One-wash 90 minute protocol
  • Sensitivity: 2.8 pg/ml
  • Range: 18.8 pg/ml - 1200 pg/ml
  • Sample type: Cell Lysate, Tissue Lysate
  • Detection method: Colorimetric
  • Assay type: Sandwich (quantitative)

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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, Tissue Lysate
  • Assay type

    Sandwich (quantitative)
  • Sensitivity

    2.8 pg/ml
  • Range

    18.8 pg/ml - 1200 pg/ml
  • Recovery

    Sample specific recovery
    Sample type Average % Range
    Fetal Bovine Serum 97 96% - 98%
  • Assay time

    1h 30m
  • Assay duration

    One step assay
  • Product overview

    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.


  • 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. 

    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.

  • Platform

    Microplate

Properties

  • Storage instructions

    Store at +4°C. Please refer to protocols.
  • Components 1 x 96 tests
    10X GFP Capture Antibody 1 x 600µl
    10X GFP Detector Antibody 1 x 600µl
    10X Wash Buffer PT (ab206977) 1 x 20ml
    4X Antibody Diluent EB 1 x 6ml
    50X Cell Extraction Enhancer Solution (ab193971) 1 x 1ml
    5X Cell Extraction Buffer PTR (ab193970) 1 x 10ml
    GFP Lyophilized Recombinant Protein 2 vials
    Plate Seals 1 unit
    Sample Diluent NS (ab193972) 1 x 12ml
    SimpleStep Pre-Coated 96-Well Microplate (ab206978) 1 unit
    Stop Solution 1 x 12ml
    TMB Development Solution 1 x 12ml
  • Research areas

    • Tags & Cell Markers
    • Fusion / Marker Proteins
    • GFP
    • Kits/ Lysates/ Other
    • Kits
    • IHC Kits
    • Kits/ Lysates/ Other
    • Kits
    • ELISA Kits
    • ELISA Kits
    • Sub cellular markers ELISA kits
    • Kits/ Lysates/ Other
    • Kits
    • ELISA Kits
    • ELISA Kits
    • Fusion/marker protein ELISA Kits
  • Relevance

    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.
  • Alternative names

    • Green Flourescent Protein

Images

  • 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

Protocols

  • Protocol Booklet

Click here to view the general protocols

Datasheets and documents

  • SDS download

  • Datasheet download

    Download

References (14)

Publishing research using ab171581? Please let us know so that we can cite the reference in this datasheet.

ab171581 has been referenced in 14 publications.

  • Yu K  et al. Transfer of oral bacteria to the fetus during late gestation. Sci Rep 11:708 (2021). PubMed: 33436911
  • Ding K  et al. Proteosomal degradation impairs transcytosis of AAV vectors from suprachoroidal space to retina. Gene Ther N/A:N/A (2021). PubMed: 33542456
  • 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
  • Wesslowski J  et al. eGFP-tagged Wnt-3a enables functional analysis of Wnt trafficking and signaling and kinetic assessment of Wnt binding to full-length Frizzled. J Biol Chem 295:8759-8774 (2020). PubMed: 32381507
  • Frederick A  et al. Engineered Capsids for Efficient Gene Delivery to the Retina and Cornea. Hum Gene Ther 31:756-774 (2020). PubMed: 32578442
View all Publications for this product

Customer reviews and Q&As

Show All Reviews Q&A
Submit a review Submit a question

1-4 of 4 Abreviews or Q&A

GFP Elisa on mice liver lysates

Excellent Good 4/5 (Ease of Use)
Abreviews
Abreviews
abreview image
I performed the GFP ELISA kit on liver lysates as detailed reported on the datasheet.
Here are the minor modifications I performed.
Livers were harvested from mice after PBS perfusion.
I homogenized 100mg of wet liver tissues in 500uL of chilled 1X cell extraction buffer PTR (prepared as reported in Abcam datasheed and supplemented with protease and phosphatase inhibitors).
Samples were minced in a tissue lyser (Qiagen, 50Hz for 5').
After homogenization samples were incubated for 20' on ice to allow proper lysis.
A subsequent centrifugation step at 4°C for 20' was used to separate unlysed fractions. Supernatants were recovered in new eppi and protein concentration is evaluated using Bradford assay.
The protocol suggested diluting samples at the desired concentration in 1X cell extraction buffer PTR. I did at 2ug, 10ug, 25ug and 50ug but in all the samples there is a brownish precipitate that makes the OD at 450nm impossible.
When I diluted 1ug of liver lysates 1:400 I finally got the ideal concentration to perform ELISA and got my result.
I had to perform some modification but I finally got it!
The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

Abcam user community

Verified customer

Submitted Mar 21 2022

Quantification of sfGFP in mouse stool

Excellent Excellent 5/5 (Ease of Use)
Abreviews
Abreviews
abreview image
Works for quantification in stool samples.
The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

Bryan Hsu

Verified customer

Submitted Dec 23 2021

Test for superfold GFP

Excellent Excellent 5/5 (Ease of Use)
Abreviews
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.
The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

Ling Chun

Verified customer

Submitted Oct 27 2014

Question

Will this work on GFP fusion proteins?

Read More

Abcam community

Verified customer

Asked on Feb 23 2016

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

Kevin Hanson

Abcam Scientific Support

Answered on Feb 23 2016

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