The anti-phospho-proteins (anti-pP) is developed using hyper-phosphorylated proteins as immunogen and affinity-purified with peptide with phosphor-serine,-threonine,and –tyrosine residues on Agarsoe. The purified antibody is immobilized to beaded agarose via strong secondary amine linkage. This product could be utilized as an affinity matrix for rapid affinity isolation, purification anf enrichment of proteins/peptides with phospho-tyosine/-serine/-threonine residues.

Immunoaffinity precipitated phosphor-proteins from milk samples using 0.5 mL anti-phosphoprotein antibody Agarose (ICP9888) and anti-biotin Agarose (ICP0615) as control.

Bound phosphoproteins were eluted with 1 mL of 0.5 M HCl with concentration of 0.6 mg/mL.

The eluted phosphoprotein was titrated with anti-phosphotyrosine antibody HRP (ICP9902).
 

Species
 

Formulation

Goat
 

0.5 mL beaded agarose suspended in 1 mL slurry

Antibody Immobilized

>10 mg/mL; antibody is covalently linked to CL-4B sepharose

The antibody selectively captured the peptides or proteins with phosphorylated Tyrosine/Serine/Threonine residues. It does not cross react with non-phosphorylated proteins.

Product is stable for several weeks at room temperature. For extended storage, store product at -20°C.  Do not aliquot and shake thoroughly before use. 

Rapid isolation, purification and enrichment of peptides or proteins with phospho-tyrosine/serine/threnonin residues from mixtures of cell lysates or protease-digested mixtures.

Protein phosphorylation is one of the most important events in post-translational modification. Protein phosphorylation has been extensively studied and is believed to be one of the switches of cell signaling. Simple and effective tools for rapid isolation and purification of the phosphorylated species is essential for proteomic profiling of the phosphorylated proteins in different stages of development, disease, and signal transduction, as well as identifying the precise site of phosphorylation in a phospho-protein.