ELISA-ONE are next generation ready to use ELISA kits for the fast detection of key intracellular proteins that undergo phosphorylation in major signaling pathways.
Protein phosphorylation plays a crucial role during intracellular signaling, but the availability of optimized and validated antibody pairs often presents a critical bottleneck for cell signaling research. ELISA-ONE kits are powered by our CaptSure technology and are comprised of precoated plates and an optimized pair of capture and detection antibodies that recognize either specific phosphoepitopes or invariant epitopes on each target.
Unlike most other ELISA protocols that require long incubation times and multiple wash steps, ELISA-ONE can be performed in no more than 90 minutes, with a single incubation and washing steps, whilst maintaining the same performance, sensitivity, and dynamic range of conventional protocols. Furthermore, the technology behind ELISA-ONE is particularly effective at eliminating the issues related to biotin interference that generally occurs in complex cell lysates.
The simplicity and efficiency of ELISA-ONE ultimately translates into faster assay setup and analysis, improved performance in complex samples and reduced cost.
The sample, the capture antibody tagged with CaptSure peptide, and the HRP-labeled detection antibody are added to the CaptSure antibody coated plate in one single step and incubated for 60 minutes. The immunocomplex formed by the analyte within the sample and the antibody pair is anchored to the plate via the interaction between the CaptSure antibody immobilized onto the plate and the CaptSure peptide conjugated to the capture antibody.
Following a washing step, unbound material is washed away and an HRP substrate (TMB) is added to the plate for colorimetric detection. The color development, due to the oxidation of TMB, is measured at 450nm and directly correlates with the amount of analyte in the sample.
The whole process takes no more than 90 minutes, cutting hands-on time in half and reducing incubation times by three times compared with conventional ELISA protocols.
ELISA-ONE kits are available for several targets, both in their phosphorylated and unphosphorylated form.
The following pathways are useful for studying key intracellular signaling events and for monitoring the activation of major receptor classes, including GPCRs, RTKs and cytokine receptors.
|AKT Signaling||STAT Signaling|
|AKT 1/2/3 (Phospho) [Ser473]||STAT3 (Phospho) [Tyr705]|
|AKT1/2/3 (Total)||STAT3 (Total)|
|AKT1/2/3 (Phospho / Total) [Ser473]||STAT3 (Phospho / Total) [Tyr705]|
|NF-κB p65 Signaling||STAT5 A/B (Phospho) [Tyr694/699]|
|NF-κB p65 (Phospho) [Ser536]||STAT5 A/B (Total)|
|NF-kB p65 (Phospho / Total) [Ser536]||STAT5 A/B (Phospho / Total) [Tyr694/699]|
|SMAD1 (Phospho) [Ser463/465]||ERK 1/2 (Phospho) [Thr202/Tyr204]|
|SMAD1 (Total)||ERK 1/2 (Total)|
|SMAD1 (Phospho / Total) [Ser463/465]||ERK 1/2 (Phospho / Total) [Thr202/Tyr204]|
All buffer and reagents required for the assay are included in the kits. If you need additional buffer packs please inquire for availability.
More ELISA-ONE kits will be available soon. However, if your target is not on the list above, find out how our CaptSure technology can enable you to use your favorite antibody pair.
ELISA-ONE kits are developed based on our CaptSure technology. CaptSure is an antibody based platform, comprising of a CaptSure antibody preimmobilized onto an ELISA assay plate that is specific for the CaptSure tag, a nonbiological peptide, covalently linked to the capture antibody of the antibody pair.
Schematic representation of CaptSure Technology.
ELISA-ONE demonstrates a wider dynamic range and lower background compared with commercially available ready to use competitor ELISA kits
Mean, standard deviation (SD) and coefficient of variance (%CV) were measured among the same plate (n=24) and four different plates