In-Cell ELISA: Benefits and Applications

In-Cell ELISA is a specific quantitative assay to measure intracellular protein concentrations with no need for cell lysis.

In-Cell ELISA is also commonly known as in-cell western, cytoblot, or cell-based ELISA. It is a useful tool that allows users to measure target analytes in situ within both suspension and adherent cells, all with minimal hands-on time and coupled with the reproducibility and specificity of an ELISA.

Abcam provides a range of In-Cell ELISA kits designed for many cell signaling proteins. It can be used to quantify cell cycle proteins, phosphorylated and non-modified proteins, as well as proteins used to spot changes in mitochondria within drug-treated cells.

Where users have existing antibodies to a specific target, Abcam is also able to offer In-Cell ELISA Support Packs that contain the full set of components and reagents required to undertake successful experiments. These Support Packs are also suitable for use with Abcam’s already well-characterized In-Cell ELISA antibodies.

Simple

  • Cell fixation to data acquisition needs around 30 minutes of hands-on time.
  • There is no need for lysate preparation or transfer.

Quantitative

  • Abcam’s product ranges include infrared (IR), fluorometric, and colorimetric detection methods. IR kits can provide improved dynamic range and sensitivity as well as allowing duplexing within wells.
  • Abcam’s products are designed to offer reliable quantitative output with HRP and IR signals able to be normalized to Janus Green staining intensity (a mitochondrial dye) to account for variation in cell numbers.
  • Archetypal intra-plate and inter-plate variability is under 10 % CV (illustrated below).

Intra-plate variability (CV) of MTCO1 is less than 10%. MTCO1 (mitochondrial cytochrome C oxidase 1) was detected using anti-MTCO1 antibody (ab14705) in HeLa cells treated with 10 µM chloramphenicol (CAM) or vehicle (DMSO) for 7 days. Sixteen datapoints for background, and 40 datapoints for DMSO and CAM were collected.

Figure 1: Intra-plate variability (CV) of MTCO1 is less than 10%. MTCO1 (mitochondrial cytochrome C oxidase 1) was detected using anti-MTCO1 antibody (ab14705) in HeLa cells treated with 10 µM chloramphenicol (CAM) or vehicle (DMSO) for 7 days. Sixteen datapoints for background, and 40 datapoints for DMSO and CAM were collected.

Inter-plate variability (CV) of MTCO1 is less than 10%. On 3 separate days, HepG2 cells were treated with 10µM chloramphenicol (CAM), 10 µM dideoxycytidine (ddC) and analyzed by In-Cell ELISA for levels of mitochondrial cytochrome C oxidase 1 (MTCO1) using anti-MTCO1 antibody (ab14705).

Figure 2: Inter-plate variability (CV) of MTCO1 is less than 10%. On 3 separate days, HepG2 cells were treated with 10µM chloramphenicol (CAM), 10 µM dideoxycytidine (ddC) and analyzed by In-Cell ELISA for levels of mitochondrial cytochrome C oxidase 1 (MTCO1) using anti-MTCO1 antibody (ab14705).

Overall, In-Cell ELISA can combine benefits from a simple technique with a quantitative output, impressive dynamic range, and well-characterized antibodies. It is an ideal assay for the detection of targets of interest within a diverse array of cell conditions.

About Abcam

Abcam is a global life sciences company providing highly validated antibodies and other binders and assays to the research and clinical communities to help advance the understanding of biology and causes of disease.

Abcam’s mission is to serve life scientists to help them achieve their mission faster by listening to their needs, continuously innovating and improving and by giving them the tools, data and experience they want. Abcam’s ambition is to become the most influential life science company for researchers worldwide.


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Last updated: Apr 1, 2019 at 6:47 AM

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