Description of Available Services

Typical Experiment Outline:

Expression Systems Available:

Purification Methods Available:

Western blotting

A strength of western blotting is the ability to investigate aspects of protein structure. The primary structural aspect is protein size.  Another important structural aspect is post-translational modifications. Many modifications will cause a change in the gel electrophoretic charateristics of the protein altering their migration patterns in the gel. These include glycosylation, phosphorylation, ubiquitination to name a few.

Western blotting allows the detection of many proteins from a single sample. This is useful for limited samples. It is very useful when analyzing an experimental pharmaceutical on cellular metabolism and cell physiology. For example, the effect of an inhibitor of growth factor signaling can be investigated by performing western blotting analysis of many proteins in multiple signaling cascades.

Western blotting is very useful in protein interaction analyses. In this case the products of protein binding assays(immuniprecipitations, GST-pulldowns) are analyzed instead of whole cell lysates. Membranes can be stored and reused.

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Plate based assays:

The ELISA is a prototypical high throughput, multi-well plate assay that is useful in research and clinical laboratories. The multi-well plate format can be adapted to a variety of protein-based and cell-based assays for quantification of protein and biomarker concentrations, protein-interactions and biological activity measurements.

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Protein interaction studies:

Protein interactions can easily be studies adapted to plate based assays. Similar to co-immunoprecipitaton antibodies are used to capture and immobilize specific protein complexes. Components of the protein-complexes are then quantified using antibody based detection methods.

Assays can be developed to identify protein-interaction partners of proteins or isolated protein interaction domains.   A specific protein can be engineered and produced as a recombinant protein fused to an epitope tags. Immobilization is accomplished by applying the fusion protein to wells coated with antibodies that recognize the epitope tag. Potential interacting partners are added  to the wells and stable protein-partner complexes detected.

Cell-based assays can be developed to monitor and quantify specific biological activities. Examples of cell-based assays are measurement of apoptosis, cell-cycle analysis.

This is a great platform for screening small molecules for protein-interaction inhibiting activity. Such as for identifying inhibitors of receptor-ligand binding interactions.

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