1. A gene’s mRNA level does not predict its protein level (PDF)
While protein levels determine cellular function and morphology, genomic approaches have dominated biomedical research, particularly since the publication of the human genome in 2004. While mRNA is required to synthesize protein, approximately 60% of the time mRNA level is not correlated to protein level.
2. Carrier Ampholine 2D SDS PAGE Overview (PDF)
Two methods of 2D electrophoresis are commonly used: IPG strips and carrier amophline (aka 2D SDS PAGE), which differ in the way the pH gradient is established and SDS compatibility. Kendrick Labs uses the 2D SDS PAGE method of 2DE, which provides the advantage of compatibility with SDS. SDS is considered the best protein solubilization reagent. 2D SDS PAGE provides a sensitive method for detection of protein by total protein staining (silver, Coomassie, etc) or via Western blotting with a specific antibody.
3. 2D SDS PAGE is useful for analysis of Host Cell Protein Antibody Coverage (PDF)
Kendrick Labs performs a 2D Western blot analysis of ELISA antibodies used for detection of host cell protein (HCP) in drug substances. This white paper provides technical details of the CA-2D SDS PAGE and Western blotting methods. The SDS compatible 2DE method we use results in a high level of protein solubility and yields sensitive, reproducible results for HCP analysis. The procedure for Western-silver image alignment and the general method of computer analysis using Progenesis SameSpots software are also described. Standard operating procedures (SOPs) regulate the entire workflow from 2DE to computer analysis.
4. Prokaryotic proteins are more easily identified than eukaryotic ones by Peptide Mass Fingerprinting (PDF)
In-house results are presented for 560 silver-stained and 830 Coomassie-stained proteins cut from 2D gels and sorted by species. Proteins for different species have varying levels of successful identification by Mass spectrometry. Sample succcess is as follows Archaea > Bacteria= Plasmidium> Mammalian> Drosophila. Results suggest that post-translational modifications on mammalian proteins are in sufficient quantity to interfere with identification by mass spectrometry, in contrast to bacterial and archaea proteins.
5. Too Much to Read (PDF)
The number of peer-reviewed publications has increased dramatically in the past decades and does not show any signs of slowing. Plotting publication numbers (1950-2016) is best represented as 5th order polynomial curve. While there are advantages in having multiple studies that support the same conclusion, it is difficult to keep current the latest research due to volume and may contribute to the lack of reproducibility of many studies.