All tags In-Cell ELISA Benefits of In-Cell ELISA

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Benefits of In-Cell ELISA

In-Cell ELISA is a quantitative method that can measure intracellular protein concentration without the need for cell lysis.

This method is also known as in-cell western, cell-based ELISA or cytoblot.  This valuable tool gives you the ability to measure target analytes in situ in both adherent and suspension cells with minimal hands on time, and with the specificity and reproducibility of an ELISA.

Abcam offers a range of In-Cell ELISA kits to many cell signalling proteins. Specifically it can be used to quantify phosphorylated and non-modified proteins, cell cycle proteins, and proteins which can be used to detect changes in mitochondria in drug-treated cells. 

If you already have your own antibodies to a specific target, we also provide In-Cell ELISA Support Packs, which contains all the reagents and components necessary to conduct successful experiments.  The Support Packs can also be used with Abcam’s well-characterized In-Cell ELISA antibodies.

​​This method is also known as In-Cell western, cell-based ELISA or cytoblot. This valuable tool gives you the ability to measure target analytes in situ in both adherent and suspension cells with minimal hands on time, with the specificity and reproducibility of an ELISA.

Our In-Cell ELISA kits detect a wide range of cell signaling proteins. They can be used to quantify phosphorylated and non-phosphorylated proteins involved in cell cycle or mitochondrial drug response.​

​​Simple - minimal hands on time

  • ​​Cell fixation to data acquisition requires approx. 30 minutes of hands on time.
  • No lysate preparation or lysate transfer is required.​

Quantitative

  • ​​​​​Our product ranges include colorimetric, fluorometric and infra-red (IR) detection methods. IR kits offer a greater dynamic range and sensitivity, and allow duplexing within wells.
  • Quantitative output - IR and HRP signals can be normalized to Janus Green staining intensity (a mitochondrial dye) to account for cell number variation. 
  • Typical intra-plate and inter-plate variability is less than 10% CV as shown in Figures 1 and 2 (respectively).
ab14705 figure 1 v2-2 in cell elisa

​​​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.  

ab14705 figure 2 v2-2

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

In summary, In-Cell ELISA combines the benefits of a simple technique with quantitative output, great dynamic range using well characterized antibodies. In-Cell ELISA is a suitable assay for detecting targets of interest in a multitude of cell conditions.

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