2010
DOI: 10.1016/j.bbrc.2010.09.104
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Detection of tyrosine phosphorylated proteins by combination of immunoaffinity enrichment, two-dimensional difference gel electrophoresis and fluorescent Western blotting

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Cited by 8 publications
(4 citation statements)
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“…Compared to Figure  2A when only the 4G10 antibody was used to visualize all pTyr proteins, fewer signals were observed when the 4G10 was combined with another antibody for detection of a specific phosphorylated protein. Imatinib treatment of K562 cells is a well characterized model system [16,17] and it was chosen since reference data was available. Data from a complementary technique is very valuable when implementing novel approaches as this paper describes.…”
Section: Resultsmentioning
confidence: 99%
“…Compared to Figure  2A when only the 4G10 antibody was used to visualize all pTyr proteins, fewer signals were observed when the 4G10 was combined with another antibody for detection of a specific phosphorylated protein. Imatinib treatment of K562 cells is a well characterized model system [16,17] and it was chosen since reference data was available. Data from a complementary technique is very valuable when implementing novel approaches as this paper describes.…”
Section: Resultsmentioning
confidence: 99%
“…Using MS‐based approaches, thousands of phosphorylations can be monitored in a sample, [ 22,23 ] but enrichment steps are needed during sample preparation to separate off high abundant native peptides that mask the detection of modified peptides. [ 24–26 ] A variety of different enrichment strategies are used, [ 24,27,28 ] and among them, metal oxides such as titanium dioxide (TiO 2 ) and zirconium dioxide are preferred due to their great tolerance to buffers and detergents. The combination of these enrichment procedures with sensitive and high‐resolution MS enables the identification and characterization of a high number of proteins and modifications simultaneously.…”
Section: Introductionmentioning
confidence: 99%
“…Some authors used chemifluorescence to reveal the antigenic map [ 55 , 63 , 64 ]. By contrast, others have used fluorescent probes for protein labeling before the electrophoresis steps [ 65 67 ]. Goeb et al [ 59 ] use a fluorescent dye for staining a landmark map in order to confirm the localization of antigenic spots as potential autoimmune targets.…”
Section: Discussionmentioning
confidence: 99%