Tyrosine kinases play a prominent role in transmitting and regulating various cell signaling pathways. Deregulation of this key PTM drives inappropriate proliferation and cell survival. MS-based global phosphoproteomics related studies have significantly evolved in recent years, but complexity of the signaling pathways and sub-stoichiometry levels of tyrosine phosphorylation sites, when compare...

Retinal S-antigen (S-ag) was purified by monoclonal antibody (MoAb) immunoaffinity chromatography from soluble protein extracts of bovine and human retina. Purification of S-ag was readily achieved by affinity chromatography using four different MoAb-Sepharose 4B columns. The four antibody columns gave different recoveries with material of comparable enrichment with greater than 95% purity as j...

new immunoaffinity fluorometric biosensor has been developed for detecting and quantifying aflatoxins, a family of potent fungi-produced carcinogens that are commonly found in a variety of agricultural products. They have also been cited as biological agents under weapons development. The handheld, self-contained biosensor is fully automatic, highly sensitive, quick, quantitative, and requires ...

A magnetic beads based immunoaffinity capillary electrophoresis method for total Immunoglobulin E quantification in serum has been developed. The method combines speed, automation ability, and minimal sample consumption. Only 1 microL of serum is required while the whole immunoaffinity capillary electrophoresis method is performed in less than 50 min. The concomitant use of online immunocapture...

A new method is described for the analysis of copper (Cu) containing ceruloplasmin (Cp) in human serum. The method uses immunoaffinity plus size-exclusion chromatography (SEC) to separate Cp from other proteins and from inorganic ions, with inductively coupled plasma mass spectrometry (ICP-MS) detection of 63Cu and 65Cu to identify the protein. A reference serum certified for Cp was used to est...

Crosstalk between the SUMO and ubiquitin pathways has recently been reported. However, no approach currently exists to determine the interrelationship between these modifications. Here, we report an optimized immunoaffinity method that permits the study of both protein ubiquitylation and SUMOylation from a single sample. This method enables the unprecedented identification of 10,388 SUMO sites ...

Factor VII has been purified to homogeneity from bovine plasma by a procedure that includes affinity purification on an immunoadsorbent column. Recovery was determined by both coagulant assay and liquid scintillation counting, using 3H-factor VII as an internal standard. The purification factor calculated by both methods was approximately 120,000-fold, with a final yield of approximately 18%. H...

Multiple approaches for simplifying the serum proteome have been described. These techniques are generally developed across different laboratories, samples, mass spectrometry platforms, and analysis tools. Hence, comparing the available schemes is impossible from the existing literature because of confounding variables. We describe a head-to-head comparison of several serum fractionation scheme...