SDS PAGE:
GBS will conduct SDS PAGE protein fractionation of protein mixtures. Clients can
choose from several total acrylamide concentrations (range 8-18%) for gels of homogenous
acrylamide percentages, or request custom gradient gels. Additionally, non-denaturing
or non-reducing PAGE can be performed. There are 3 different size formats to choose
from, and several different total and specialized protein staining methods. Optional
quantitative and qualitative analysis is also available.
For details on SDS PAGE Services,
click here.
Rapid Analysis Meso-Resolution™ (RAMR™) Protein Profiling:
RAMR protein profiling is a novel proteomic approach developed by GBS that provides
medium resolution protein profiling in timelines of less than half of that required
for 2DE. The procedure typically resolves 120-150 proteins and is 4 to 5 times as
sensitive as standard SDS PAGE in detecting changes in protein abundance. In addition,
customized chromatographic prefractionations followed by SDS PAGE analysis capable
of focused higher resolution is available upon request.
For details on RAMR™ Services,
click here.
2-D Electrophoresis:
GBS will perform two-dimensional electrophoresis (2DE) on sample materials. The
first dimension is conducted with immobilized pH gradient (IPG) strips using a Amersham
Biosciences Multiphor II system. The system used for the second dimension depends
on which size gel format is required. You can submit sample materials ready for
analysis, or raw materials including tissues, tissue cultures, cell lines, intact
bacteria, yeast and other microorganisms and let GBS process the raw material. If
you wish to conduct initial sample disruption inhouse, GBS can mail its optimized
denaturing buffer cocktail for final sample dilution. For isoelectric focusing (first
dimension) customers may choose from a variety of IPG strips of broad to very narrow
pH ranges, and from up to 4 strip lengths. For the second dimension, the customer
may choose from several total acrylamide concentrations (range 8-18%) for gels with
homogenous acrylamide percentages, or request custom gradient gels. Additionally
customers may request 2DE under nondenaturing or nonreducing conditions in either
dimension or both. There are 3 different size formats to choose from, and several
different total and specialized protein staining methods.
For details on 2-D Electrophoresis,
click here.
Differential Proteomic Analysis
After 2DE separation, GBS can perform sample analysis using Phoretix 2D Evolution
software (Nonlinear Dynamics). Minimal analysis includes spot detection, calculation
of estimated molecular weights, a normalized quantitative value and pI of each spot.
Advanced analysis also includes spot matching between different gels and differential
comparison of equivalent spot abundance values. For experiments with multiple replicates
per experimental group averaged gels are synthesized and used for differential analysis.
When multiple experimental groups are present, each experimental group is compared
to a reference gel synthesized from the control gel(s).
For details on Differential Proteomic Analysis,
click here.
Hi-Res Proteomic Analysis using Prefractionation
GBS can use its optimized chromatographic prefractionation (used in RAMR protein
profiling procedure) but apply 2DE rather than SDS PAGE to the isolated fractions
providing the enhanced proteomic analysis. This procedure results in three rather
than one 2DE gels per sample. There is relative small overlap between the different
fractions so the enrichment effects of the prefractionation allow enhanced sensitivity
and resolution compared to standard 2DE and other prefractionation procedures. Additionally,
custom chromatographic prefractionations prior to 2DE analysis can be developed
in conjunction with GBS staff.
For details on Hi-Res Proteomic Analysis using Prefractionation,
click here.