With regard to 2D-PAGE
conditions, before 2D-PAGE
, the strip gel was equilibrated in sample buffer (strip equilibration buffer; 6 M urea, 20% glycerol, 2% dithiothreitol (DTT), 2% sodium dodecyl sulfate (SDS), and 375 mM Tris-HCI, pH 8.
162 proteins distribute among pI 4-10 intervals but 21 proteins with pI >10 were identified that beyond the 2D-PAGE
CFTR-enriched endosomes were visualized by immuno-EM, affinity purified by AminoLink[R] gel, and analyzed by 2D-PAGE
Except the outcome isn't quite so rewarding when all us journos are trying to do is comb through a 2D-page
submission from Costcutter.
The following chapter provides an excellent discussion of quantitative proteomic techniques, covering 2D-PAGE
techniques, multiplexing, metabolic and postextraction labeling, and isolation and quantitation of phosphopeptides.
Although various approaches have been developed for protein expression analysis (4), the combination of 2D-PAGE
and mass spectrometry (MS) has been the most successful (4-7).
Our functional genomic investigations will include quantitative measurements of mRNA using expressed sequence tags (ESTs) and microarrays, protein expression using 2D-PAGE
and mass spectrometry, and metabolite expression using various analytical platforms to gain a multi dimensional view of cellular functions and biology," notes Dr.
In addition to the existing products for improved 2D-PAGE
sample preparation, Vivascience is now also offering spin columns for sample preparation prior to mass spectrometry.
Convenient and reproducible sample preparation prior to 2D-PAGE
1] Although 2D-PAGE
is unchallenged in its ability to resolve thousands of proteins, it is labor-intensive, requires large quantities of protein, and is not easily converted into a diagnostic test.
Proteins separated by 2D-PAGE
were identified by mass-mapping studies (see below).
Although such an approach represents the gold standard for global analysis of protein expression, 2D-PAGE
often is impractical for diagnostic applications.