Overview

  • Product nameIn-Cell ELISA (ICE) Support Pack
    See all In cell elisa kits
  • Sample type
    Adherent cells, Suspension cells
  • Assay typeCell-based (quantitative)
  • Assay durationMultiple steps standard assay
  • Product overview

    ab111542 is for use with suspension, apoptotic/detaching cells and adherent cell lines. The pack contains five 96-well microplates, buffers and protocol to perform ICE with Abcam's ICE-validated antibodies. For an ICE assay, it is necessary to purchase both primary antibody(ies) and labeled secondary antibody(ies). Antibodies are sold separately, allowing customizing the target(s) of interest, method of detection and multiplexing.


     

    In-Cell ELISA uses quantitative immunocytochemistry to measure protein levels or post-translational modifications within cells. The cells are fixed to the bottom of a coated 96-well plate (provided). Targets of interest are detected by primary antibodies, which are in turn quantified with labeled secondary antibody(ies). Abcam offers highly-specific, well-characterized primary antibodies, IRDye®- or HRP-labeled anti-mouse and anti-rabbit secondary antibodies, as well as IRDye®-labeled isotype specific anti-mouse antibodies. By combining antibodies of different species or isotype and appropriate IR-labeled secondary antibodies, two color multiplexing can be achieved in the 800/700 channels. IR imaging and quantification is performed using a LI-COR® Odyssey® or Aerius® system. HRP-labeled complexes are developed and quantified colorimerically using spectrophotometer.


    Two protocols are available when using this kit. Protocol (1) suspension cells and cells likely to detach under experimental conditions (for example, adherent cells undergoing apoptosis readily detach from a culture plate). Protocol (2) - A second protocol is available for normal adherent cells. These protocols can be used with any of Abcam's ICE-validated antibodies. Specific scientific information, background and working concentration for each antibody are detailed in each antibody's corresponding datasheet.

     

  • Tested applicationsSuitable for: In-Cell ELISAmore details
  • PlatformMicroplate

Properties

  • Storage instructionsStore at +4°C. Please refer to protocols.
  • Components 5 x 96 tests
    100X Triton X-100 1 x 1.25ml
    10X Blocking Solution 1 x 40ml
    10X Phosphate Buffered Saline (PBS) 1 x 250ml
    400X Tween-20 1 x 4ml
    96-well Assay Plate 5 units
    Janus Green Stain 1 x 30ml
    Plate Seals 5 units

    Applications

    Our Abpromise guarantee covers the use of ab111542 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
    In-Cell ELISA Use at an assay dependent dilution.

    In-Cell ELISA (ICE) Support Pack images

    • Cells are grown to ~80% confluency in a 96- or 384-well plate, a drug/other treatment is applied to stimulate a cellular response. The cells are then fixed and permeabilized, effectively "freezing" them. Primary antibodies are then added which bind to their intended targets within the mitochondria or other subcellular compartment. After incubation, the unbound primary antibodies are washed away and secondary antibodies are added. These secondaries are conjugated to either IRDyes® or to an enzyme label (HRP or AP) for the colorimetric versions of the assays. Unbound secondaries are washed away, reaction buffer is added for the colorimetric assays, and the signal is read on a suitable instrument for the kit type.
      » In-cell ELISA diagram in PDF format

    Protocols

    References for In-Cell ELISA (ICE) Support Pack (ab111542)

    ab111542 has not yet been referenced specifically in any publications.

    Product Wall


    1. ab126420, the components of the blocking buffer are proprietary, and the solution is a standard blocking solution.

    2. ab111542: The blocking solution in the kit is the same as the 10X blocking solution sold separately (ab126587). It is...

    Read More

    Thank you for your inquiry.

    I am happy to confirm that ICE can be used for nuclear proteins without any deviations from the standard protocol.

    Please have a look at this link where more information can be found:

    http://www...

    Read More

    I am very pleased to hear you would like to accept our offer and testab616 and ab15580in ICE. These codes will give you: 1 free primary antibody per code before the expiration date. To redeem this offer, please submit an Abreview for ICE for each antib...

    Read More

    Thank you very much for your feedback! We are happy to help answer any technical questions you may have about In-Cell ELISA or any of our other assay or products. Please let me know if you need any additional information or assistance.

    Thank you for contacting us. Yes, In-Cell ELISA can be performed on suspension cells. Our ICE Support Pack ab111542 has been optimized for use on both adherent and suspension cells and provides the attached protocol for performing ICE on Suspension Cel...

    Read More

    Thank you for your inquiry.



    Unfortunately, we don’t offer the MetAbArray service any longersince we are 100% dedicated to individual product/assay development.


    All of the antibodies that went into that offerin...

    Read More

    Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"