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.