2016
DOI: 10.1074/mcp.m116.059311
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A Human Lectin Microarray for Sperm Surface Glycosylation Analysis

Abstract: Glycosylation is one of the most abundant and functionally important protein post-translational modifications. As such, technology for efficient glycosylation analysis is in high demand. Lectin microarrays are a powerful tool for such investigations and have been successfully applied for a variety of glycobiological studies. However, most of the current lectin microarrays are primarily constructed from plant lectins, which are not well suited for studies of human glycosylation because of the extreme complexity… Show more

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Cited by 26 publications
(15 citation statements)
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“…Washed RBC were subjected to ice cold hypotonic lysis in 20 mM Tris, pH 7.6 and protease inhibitor (05056489001, Roche) (31). Lysates were washed three times in hypotonic lysis buffer (37000 g, 4°C, 30 minutes) before resuspension in minimal hypotonic lysis buffer.…”
Section: Rbc Ghost Preparationmentioning
confidence: 99%
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“…Washed RBC were subjected to ice cold hypotonic lysis in 20 mM Tris, pH 7.6 and protease inhibitor (05056489001, Roche) (31). Lysates were washed three times in hypotonic lysis buffer (37000 g, 4°C, 30 minutes) before resuspension in minimal hypotonic lysis buffer.…”
Section: Rbc Ghost Preparationmentioning
confidence: 99%
“…Ghost preparations were mixed in equal volumes with SDS sample buffer containing 8M urea (31) and heated at 100°C for 10 minutes. Ghost protein samples were fractionated by gel electrophoresis using…”
Section: Lectin Western Blotmentioning
confidence: 99%
See 1 more Smart Citation
“…Huang et al, 2004; Kung et al, 2009). Importantly, because they are also amenable to activity assays designed to determine enzyme-substrate relationships, functional protein microarrays have been particularly useful for determining the global substrate specificity of key signaling enzymes(Cox et al, 2015; Jeong, Rho, & Zhu, 2011; Lu, Lin, Boeke, & Zhu, 2013; Sun et al, 2016). For example, to gain a better understanding of the organization of human phosphorylation networks, we recently examined the global substrate profiles of 289 unique human kinases using functional protein microarrays composed of ~4,200 full-length human proteins (Newman et al, 2013).…”
Section: Section 2 Strategies To Characterize Global Changes In the mentioning
confidence: 99%
“…This information will be particularly useful for understanding the functional consequences of phosphorylation and for developing new tools, such as phospho-specific antibodies and novel FRET-based kinase activity reporters, used to study dynamic phosphorylation networks. Moreover, the generalizability of this approach suggests that it will be useful for examining the global substrate profiles of enzymes involved in SUMOylation (Cox et al, 2015), ubiquitylation (Jeong et al, 2011), acetylation (Lu et al, 2013) and O-glycosylation (Sun et al, 2016), which together will offer clues about potential points of intersection between different signaling modalities.…”
Section: Section 2 Strategies To Characterize Global Changes In the mentioning
confidence: 99%