Product nameIn-Cell ELISA (ICE) Support Pack
See all In cell elisa kits
Sample typeAdherent cells, Suspension cells
Assay typeCell-based (quantitative)
Assay durationMultiple steps standard assay
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.
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
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
This product has been referenced in:
- Siqueira RAGB et al. When spider and snake get along: Fusion of a snake disintegrin with a spider phospholipase D to explore their synergistic effects on a tumor cell. Toxicon 168:40-48 (2019). Read more (PubMed: 31251993) »
- Wang RE et al. Rational design of a Kv1.3 channel-blocking antibody as a selective immunosuppressant. Proc Natl Acad Sci U S A 113:11501-11506 (2016). Read more (PubMed: 27663736) »