GFP ELISA Kit (ab171581)
- Datasheet
- References (3)
- Protocols
Overview
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Product nameGFP ELISA Kit
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Detection methodColorimetric
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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 typeCell Lysate, Cell culture media, Tissue Lysate
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Assay typeSandwich (quantitative)
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Sensitivity1.8 pg/ml
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Range2.7 pg/ml - 2000 pg/ml
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Recovery
Sample specific recovery Sample type Average % Range Cell culture media 102 101% - 103% Fetal Bovine Serum 97 96% - 98% -
Assay time1h 30m
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Assay durationOne step assay
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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.
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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.
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Tested applicationsSuitable for: Sandwich ELISAmore details
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PlatformMicroplate
Properties
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Storage instructionsStore at +4°C. Please refer to protocols.
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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 1 x 12ml SimpleStep Pre-Coated 96-Well Microplate (ab206978) 1 x 96 tests Stop Solution 1 x 12ml TMB Development Solution 1 x 12ml -
Research areas
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RelevanceEnergy-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.
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Alternative names
- Green Flourescent Protein
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
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Example GFP standard curveBackground-subtracted data values (mean +/- SD) are graphed.
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GFP cross reactivity dataThis kit recognizes both GFP and enhanced GFP (eGFP) in cell and tissue extracts. No reactivity with mCherry, RFP or YFP was observed.
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Titration of GFP spiked 143B cell lysate within the working range of the assayBackground subtracted data from triplicate measurements are plotted.
Datasheets and documents
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) »
