Outline

The Flinders Protoeomic Facility (FPF) was established in October, 2002, with funds from the NHMRC, FMRI and the Ramaciotti Foundation.

The Flinders Protoeomic Facility provides the Flinders Medical Centre, Flinders University and other South Australian institutions with state of the art equipment and expertise in the Proteomics field.

Proteomics in its broadest definition is the study of proteins. Most commonly, it uses 2-dimensional electrophoresis for the separation of protein extracts and mass spectrometry to identify the proteins in these large sets of proteins. However, these techniques can also be applied to the study of individual proteins, particularly in the analysis of post-translational modifications.

The main focus of the FPF is to provide equipment, expertise and training in 1D and 2D SDS PAGE and 1D and 2D Western blots.

The FPF also supports other protein techniques including immunoprecipitation, Liquid Chromatography, HPLC of proteins, protein quantification assay, protein purification and organelle purification.

Location

The FPF is located in Laboratory 6E404 of the Flinders Medical Centre.

Equipment

2D-Electrophoresis

1st dimension

7 cm, 11 cm, 13 cm, 18 and 24 cm isoelectric focussing with 2 IPGPhor isoelectric focussing units.

 

2nd dimension

Mini-gel (8 x 10 cm), medium format (16 x 16 cm) or large format (Ettan) (18 x 24 cm) 1D and 2D SDS PAGE gels.

 


2D-Western blots

Mini and medium format 2D Western blots.

Imaging

An Amersham VDS-CL is available to image most types of gels and Western blots. The table below lists the

Protein Western blot DNA
Sypro Ruby ECF Ethidium Bromide
Silver stain ECL  
Coomassie blue ECL Plus  
Deep Purple ECL Advanced  

Spot Cutting

A Onetouch 2D gel spot cutter with disposable tips is available to cut spots for mass spectrometry.

Mass Spectrometry

Identification of proteins can be performed using a Micromass MALDI-TOF mass spectrometry. Target proteins are excised from the gel, digested with Trypsin and the digested protein spotted onto a MALDI target plate ready for mass spectrometry. The mass spectrometer is housed in the Physical Sciences building of Flinders University and is managed by Dr Daniel Jardine (http://www.bioinnovationsa.com.au/aib_labs/site_view.php?site_id=2).

Data Analysis

A workstation is connected to the mass spectrometry facility with Micromass software for the analysis of mass spectrometry data. Analysis of 2D gels can also be performed using a number of software packages.

Costs

Performing your own experiments
Users of the FPF are required to replace all consumables that they use including reagents, buffers and disposable laboratory items.

Having your experiments run for you
A service is available whereby your 2D gels, Western blots and mass spectrometry can be performed for you. For prices, please contact Dr Tim Chataway.

Further Information

For further information, contact
Dr Tim Chataway
Lab 6E404 (phone 64996) or 5E115.1 (phone 65608)
Email: tim.chataway@flinders.edu.au