mCherry ELISA Kit (ab221829)
Key features and details
- One-wash 90 minute protocol
- Sensitivity: 0.47 pg/ml
- Range: 6.25 pg/ml - 400 pg/ml
- Sample type: Cell culture extracts, Cell culture media, Cell culture supernatant
- Detection method: Colorimetric
- Assay type: Sandwich (quantitative)
Overview
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Product name
mCherry ELISA Kit
See all mCherry kits -
Detection method
Colorimetric -
Precision
Intra-assay Sample n Mean SD CV% Media 8 2.9% Inter-assay Sample n Mean SD CV% Media 3 3.7% -
Sample type
Cell culture supernatant, Cell culture extracts, Cell culture media -
Assay type
Sandwich (quantitative) -
Sensitivity
0.47 pg/ml -
Range
6.25 pg/ml - 400 pg/ml -
Recovery
Sample specific recovery Sample type Average % Range Cell culture supernatant 111 110% - 113% Cell culture extracts 100 97% - 102% Cell culture media 112 111% - 112% -
Assay time
1h 30m -
Assay duration
One step assay -
Product overview
mCherry ELISA Kit (ab221829) is a single-wash 90 min sandwich ELISA designed for the quantitative measurement of mCherry protein in cell culture extracts, cell culture media, and cell culture supernatant. It uses our proprietary SimpleStep ELISA® technology. Quantitate mCherry with 0.47 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 stripsA 384-well SimpleStep ELISA® microplate (ab203359) is available to use as an alternative to the 96-well microplate provided with SimpleStep ELISA® kits.
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Notes
This kit recognizes mCherry fluorescent protein from the organism Anaplasma marginale. mCherry is a general-purpose, red monomer, fluorescent protein with superior photostability. The mCherry fluorescent protein is a variant of the Discosoma red (DsRed) protein and is excited at wavelengths longer than 600 nm.
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 (12 x 8 well strips)
Properties
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Storage instructions
Store at +4°C. Please refer to protocols. -
Components 1 x 96 tests 10X mCherry Capture Antibody 1 x 600µl 10X mCherry Detector Antibody 1 x 600µl 10X Wash Buffer PT (ab206977) 1 x 20ml 50X Cell Extraction Enhancer Solution (ab193971) 1 x 1ml 5X Cell Extraction Buffer PTR (ab193970) 1 x 10ml Antibody Diluent 4BC 1 x 6ml mCherry Lyophilized Recombinant Protein 2 vials Plate Seals 1 unit Sample Diluent NS (ab193972) 1 x 50ml SimpleStep Pre-Coated 96-Well Microplate (ab206978) 1 unit Stop Solution 1 x 12ml TMB Development Solution 1 x 12ml -
Research areas
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Relevance
mCherry is derived from proteins originally isolated from Cnidarians (jelly fish, sea anemones and corals), and is used as a fluorescent tracer in trasfection and transgenic experiments. The prototype for these fluorescent proteins is Green Fluorescent Protein (GFP), which is a ~27kDa protein isolated originally from the jellyfish Aequoria victoria. The mCherry protein is derived from DsRed, a red fluorescent protein related to GFP isolated from so-called disc corals of the genus Discosoma.
Images
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Other - mCherry ELISA Kit (ab221829)
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.
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Example of mCherry standard curve in Sample Diluent NS.Background-subtracted data values (mean +/- SD) are graphed.
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Example of mCherry standard curve in 1X Cell Extraction Buffer PTR.Background-subtracted data values (mean +/- SD) are graphed.
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Interpolated concentrations of spiked mCherry in cell culture supernatant samples.The concentrations of mCherry were measured in duplicates, interpolated from the mCherry standard curves and corrected for sample dilution. Undiluted samples are as follows: RPMI culture media with 10% FBS (100%), and HUVEC supernatant (100%). The interpolated dilution factor corrected values are plotted (mean +/- SD, n=2). The mean mCherry concentration was determined to be 204.81 pg/mL in spiked RPMI culture media with 10% FBS (100%) and 212.94 pg/mL in spiked HUVEC supernatant (100%). HUVEC cells were cultured in RPMI media with 10% FBS and supernatant collected.
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Interpolated concentrations of spiked mCherry in HEK 293T and 143B (wild-type) cell extract samples based on a 200 µg/mL extract load.The concentrations of mCherry were measured in duplicate and interpolated from the mCherry standard curve and corrected for sample dilution. The interpolated dilution factor corrected values are plotted (mean +/- SD, n=2). The mean mCherry concentration was determined to be 182.08 pg/mL in spiked HEK 293T cell extract and 198.36 pg/mL in spiked 143B cell extract.
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Raw O.D. values are shown for mCherry protein, YFP, GFP, and RFP.All proteins were loaded at 400 pg/mL. The average background O.D. is represented by the dashed line. Signal from YFP, GFP, and RFP loaded at 400 pg/mL was equivalent to background.
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Sandwich ELISA - mCherry ELISA Kit (ab221829)To learn more about the advantages of recombinant antibodies see here.
