Detection of signal pathway phosphorylation level by Chip

Radio-tagged studies have shown that about 30% of proteins can be phosphorylated in eukaryotic cells. Reversible phosphorylation results in changes in protein activity and function. This critical post-translational modification regulates a wide range of cellular activities, thereby affecting and regulating cellular functions such as energy storage, cellular morphological changes, protein synthesis and gene expression, release of signaling factors, muscle contraction, and cellular metabolism.

Common tests for phosphorylation levels include phospho-specific antibody, kinase activity assays, western blot, enzyme-linked immunosorbent assay (ELISA), cell-based ELISA, intracellular flow cytometry and ICC/IHC, mass spectrometry and protein Chip. Protein microarray is widely used because of its high throughput and multiple coverage.

Phosphorylated antibody chip is mainly based on the glass slide. After chemical modification on the chip, antibodies are placed on the chip by using the patented technology of three-dimensional polymer membrane. In this way, the 679 phosphorylation sites of 432 signaling proteins can be detected by high density combination of 1318 highly specific antibodies on the chip substrate. These signaling proteins are widely involved in signal transduction in many important signaling pathways. Moreover, a pair of antibodies are set to detect the phosphorylation and non-phosphorylation states of each specific protein phosphorylation site to improve the sensitivity and stability of phosphorylation detection, so as to achieve comprehensive screening of multiple signaling pathways.

Phosphorylated antibody chip features:

• 1×25×76mm silicon wafer, protein point diameter ~60μm, volume 0.5-1nL;
• Technical repeats: 6 technical repeats for each antibody site;
• Fluorescence detection: single or multi-color labels can be used;
• Suitable for tissue, cell and other samples;
• Purification system: All proteins were expressed and purified by eukaryotic yeast expression system under non-denaturing conditions.
• Production conditions: temperature 4-8℃, humidity 30-40% sample, fixed overnight at 4℃, long-term storage at -80℃.

We provide phosphorylated antibody chips including but not limited to the following signaling pathways:

• AKT pathway phosphorylation antibody chip
• Cytoskeleton signaling pathway phosphorylated antibody microarray
• CREB signaling pathway phosphorylated antibody chip
• Cell Cycle Phosphorylated antibody chip
• GPCR related protein phosphorylation antibody chip

Above all, we have established a complete phosphorylated antibody chips platform to provide testing service assistance according to customer needs.

Workflow of phosphorylated antibody chips testing service

If you are interested in our phosphorylated antibody chips service, please feel free to contact us. Profacgen looks forward to cooperating with you.