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, 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).
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.
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.