pIMAGO-Biotin for Western Blot HRP Based Detection (ECL)
pIMAGO is a universal phosphoprotein detection technology, that is a reliable alternative to phosphoantibodies for Western blot or ELISA, and a safe and cost effective alternative to 32P radioactive assays. It enables sensitive and specific recognition of phosphorylated molecules.
Unlike phosphoantibodies, the binding is not biased by amino acid sequence, and therefore can be used for detection of any phosphorylation event on any protein site. The pIMAGO detection protocol resembles a simple Western blot procedure and is easy to incorporate into any laboratory.
- Universal detection – Detection in any organism / sequence / amino acid through Ti4+
- High specificity – Multiple Ti4+ ensure strong binding to resist harsh washing
- High sensitivity – Multiple biotin ensures significant signal amplification (100pg)
- Multiplex capability – Small size (4nm) allow multiplexing with total protein antibodies
- Flexibility – Avidin–HRP / Fluor 550 / Fluor 680 / Fluor 800 available for detection.
Results and Data:
pIMAGO is based on water soluble, globular nanopolymers (i.e., dendrimers) that are multifunctionalized with reactive groups (e.g., Ti(IV) ions) for the highly specific recognition of phosphate groups, and with “reporting” groups (e.g., biotin, fluorophore) that facilitate their detection. Multiple functionalized groups per dendrimer ensure very strong binding to the phosphoproteins (resulting in high specificity) and significant signal amplification from many “reporting” groups (resulting in high sensitivity).
pIMAGO Analysis of Six Protein Mixture (Two Phosphoproteins) Avidin–HRP or Avidin–Fluor Detection.
|Avidin–HRP Detection||Avidin–Fluor680 Detection|
|Amount of protein mix loaded (in ng)||Amount of protein mix loaded (in ng)|
pIMAGO Analysis of In Vitro Kinase Assays using Avidin–HRP (left) or Avidin–Fluor (right) Detection.
|JNK1–cJun Kinase Assay||Cdk1–Rb Kinase Assay|
|pIMAGO α-cJun Overlay||pIMAGO α-cJun Overlay|
Cdk1–Cdc6 Kinase Assay
- Phosphorylation detection by Western blot or dot-blot
- Phosphorylation detection by ELISA, microplate assays
- In vitro kinase / phosphatase assays
- Kinase activity profiling / inhibitor screening
- Detection of in vivo phosphorylation after IP
- Phospho array currently being investigated.