Scientists have long used a combination of two-dimensional gel electrophoresis to study protein expression; mass spectrometry to identify unknown proteins; and enzyme-linked immunosorbent assays to watch protein activity and interaction. The results can take weeks. A number of start-up firms believe ...
AD, and CBD; the sample amount added per lane were 5 µg and 1 µg, respectively. Blotting was performed using PHF-1 mAb detecting tau phosphorylated at Ser396/Ser404 (courtesy of Dr. Peter Davies). Samples were reduced in electrophoresis sample buffer containing 2.5% β...
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Proteome analysis of hepatocellular carcinoma by two-dimensional difference gel electrophoresis: novel protein markers in hepatocellular carcinoma tissues. Mol Cell Proteomics. 2007;6:1798–808. CAS PubMed Google Scholar Yang MH, Tyan YC, Jong SB, Huang YF, Liao PC, Wang MC. Identification of ...
variety of conventional methods including: liquid chromatography such as normal or reversed phase, using HPLC, FPLC and the like; affinity chromatography (such as with inorganic ligands or antibodies); size exclusion chromatography; immobilized metal chelate chromatography; gel electrophoresis; and the ...
Cell lysate or sucrose gradient fractions were loaded in 4–12% precasted SDS-polyacrylamide gel electrophoresis gradient gels, and the resolved proteins were transferred to Hybond C nitrocellulose membranes (GE Healthcare). The membranes were blocked in 5% nonfat milk in Tris-buffered saline ...
After electrophoresis, SDS polyacrylamide gels were stained with G250 Coomassie blue. GST-MZF1 was excised and digested in-gel with sequencing-grade, modified trypsin (Promega). In brief, the GST-MZF1 spot was excised from the gel, placed in a polypropylene tube, and washed four to five ...
Electrophoresis cannot account for the translocation as the force at negative potentials would drive the positively charged trypsin at pH 8.0 (and GFP at pH 5.0) out of the pore (Supplementary Fig. 12). Most likely, the details of molecular transport must also consider the non-simplistic ...
Using free flow electrophoresis (FFE), multiple IGFBP-1 phosphoisoforms in amniotic fluid were resolved within pH 4.43–5.09. The binding of IGFBP-1 for IGF-I in each FFE fraction was determined with BIAcore biosensor analysis. The IGF-I affinity (...