概述

  • 产品名称
    mCherry ELISA试剂盒
  • 检测方法
    Colorimetric
  • 精确度
    批次内
    样品 n Mean SD CV%
    Media 8 2.9%
    批次间
    样品 n Mean SD CV%
    Media 3 3.7%
  • 样品类型
    Cell culture supernatant, Cell culture extracts, Cell culture media
  • 检测类型
    Sandwich (quantitative)
  • 灵敏度
    0.47 pg/ml
  • 范围
    6.25 pg/ml - 400 pg/ml
  • 复检

    特定样本复检结果
    样品类型 平均% 范围
    Cell culture supernatant 111 110% - 113%
    Cell culture extracts 100 97% - 102%
    Cell culture media 112 111% - 112%

  • 检测时间
    1h 30m
  • 实验步骤
    One step assay
  • 产品概述

    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


     

  • 说明

    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.

  • 经测试应用
    适用于: Sandwich ELISAmore details
  • 平台
    Microplate (12 x 8 well strips)

性能

  • 存放说明
    Store at +4°C. Please refer to protocols.
  • 组件 1 x 96 tests
    10X mCherry Capture Antibody 1 x 600µl
    10X mCherry Detector Antibody 1 x 600µl
    mCherry Lyophilized Recombinant Protein 2 vials
    Antibody Diluent 4BC 1 x 6ml
    10X Wash Buffer PT (ab206977) 1 x 20ml
    5X Cell Extraction Buffer PTR (ab193970) 1 x 10ml
    50X Cell Extraction Enhancer Solution (ab193971) 1 x 1ml
    TMB Substrate 1 x 12ml
    Stop Solution 1 x 12ml
    Sample Diluent NS 1 x 50ml
    SimpleStep Pre-Coated 96-Well Microplate (ab206978) 1 unit
    Plate Seals 1 unit
  • 研究领域
  • 相关性
    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.

应用

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.

应用 Ab评论 说明
Sandwich ELISA Use at an assay dependent concentration.

图片

  • 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.

实验方案

文献

ab221829 has not yet been referenced specifically in any publications.

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