Bio-Rad 2D Gel Electrophoresis System

A complete Bio-Rad 2D gel system is available for isoelectric focusing and SDS-PAGE protein fractionations.  Gel casting and equipment for running up to 12 samples at once is available.  The equipment includes the following:

Protean IEF, Protean Plus Dodecacell, Protean Plus Multicaster, Glass plates, Combs and Gel Stand, Focusing tray and lid, Rehydration/Equilibration tray

LSBI Contact: Dr. Tibor Pechan

Proxeon Nano 2D LC-iTRAQ

Our X'TremeSimple Splitless NanoLC system is configured to separate iTRAQ-labeled protein digests by 2D LC followed by direct spotting to MALDI targets for MS/MS analysis.  Four high precision pumps are set to produce two binary gradients.  The first binary gradient, water and acetonitrile, spots fractions from the RP C18 column.  The second binary gradient, water and ammonium acetate, controls elution from the SCX column onto the RP column.  A 10-port time-programmable switching valve connects the two RP columns in parallel to one SCX column.  While fractions from one RP column are being spotted, the other RP column collects eluent from the SCX column, then is desalted prior to the next reverse phase gradient.

Product Bulletin | iTRAQ Chemistry Guide | Chemistry Reference Card | Application Note - Yeast Example | Ross et al 2004 | Tech Note on Quantification | iTRAQ GPS | Matrix Reduction Application Notes | Non-redundant RDA Analysis | Protein-Protein Interaction, Affinity Pulldowns

LSBI Contact: Dr. Tibor Pechan

Bio-Rad Robotic Proteome Works Spot Cutter

Following the staining of protein spots, the Bio-Rad Spot Cutter/Imager integrates spot cutting with PDQuest image analysis software to cut protein spots from polyacrylamide gels or PVDF membranes.  This system can accommodate any hand cast or commercial gels (backed or free) or membranes up to 26 x 24 cm.  This system reduces manual labor/error and improves throughput of protein spots for analysis.  The PDQuest analysis system can resolve overlapped spots and distinctly target them for cutting, separating them to reduce the impact of protein mixtures in identification analyses.  Spot cutting lists can be exported to a spreadsheet for tracking data with any type of protein identification method.

LSBI Contact: Dr. Tibor Pechan

Genomic Solutions Robotic Digester & Spotter

LSBI Contact: Dr. Tibor Pechan

Genomic Solutions Investigator ProPrep 4 Block System

The Investigator ProPrep is a fully integrated system for automated in-gel enzymatic digestion of proteins and spotting for MALDI mass spectrometry.  Protocols for all steps of in-gel digestion (washing, reduction, alkylation and proteolytic digestion) are preprogrammed and may be modified as desired.  In addition the instrument automates sample cleanup procedures of the peptide fragment samples, addition of matrix, and spotting onto MALDI plates.  The peptide fragment samples can also be replated into 96 or 384 well plates for analysis by other methods such as LC/MS/MS.

LSBI Contact: Dr. Tibor Pechan

GE HealthCare 2-D Fluorescence Difference in Gel Electrophresis (Ettan DIGE)

The Ettan DIGE system allows for the comparative analysis of two or three protein samples that have been mixed together and fractionated on a single gel by 2D gel electrophoresis.  This is accomplished by labeling the samples with different fluorescent CyDyes.  This approach eliminates gel-gel variation and provides a method to use the same internal control on every gel.  The Ettan DIGE system consists of the Amersham Biosciences proprietary cyaninedye chemistry (CyDye DIGE fluors), the Typhoon gel imager, and the DeCyder differential analysis software.

User's Manual

LSBI Contact: Dr. Tibor Pechan

GE HealthCare Robotic Ettan Spot Picker

The Ettan Spot Picker is designed to work directly with the DeCyder 2-D gels analysis software enabling picking of the spots/proteins of interests.  Although the set is capable of handling gels stained by array of different dyes, at the LSBI, the Ettan Spot Picker is mainly used for the DIGE (Difference in gel electrophoresis) experiments utilizing CyDyes (User's Manual).

LSBI Contact: Dr. Tibor Pechan