Overview

  • 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 in vitro SimpleStep ELISA® (Enzyme-Linked Immunosorbent Assay) kit is designed for the quantitative measurement of mCherry protein in cell culture supernatant and cell extract samples.


    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.


    Sensitivity:
    Samples diluted in Sample Diluent NS – 0.47 pg/mL
    Samples diluted in 1X Cell Extraction Buffer PTR – 3.37 pg/mL


     

  • 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.

  • Tested applications

    Suitable for: Sandwich ELISAmore details
  • Platform

    Microplate (12 x 8 well strips)

Properties

  • 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 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

  • 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.

Associated products

Applications

Our Abpromise guarantee covers the use of ab221829 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

  • 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.

     

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

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

  • 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.

  • 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.

  • 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.

Protocols

References

This product has been referenced in:

  • 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). Read more (PubMed: 30082848) »
See 1 Publication for this product

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