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LIBRIS Formathandbok  (Information om MARC21)
FältnamnIndikatorerMetadata
00003163nam a2200325 4500
001oai:DiVA.org:uu-236588
003SwePub
008141120| | |||||||||||000 ||eng|
0247 a https://urn.kb.se/resolve?urn=urn:nbn:se:uu:diva-2365882 URI
040 a (SwePub)uu
041 a engb eng
042 9 SwePub
072 7a vet2 swepub-contenttype
072 7a ovr2 swepub-publicationtype
1001 a Yan, Junhongu Uppsala universitet,Institutionen för immunologi, genetik och patologi4 aut0 (Swepub:uu)Junya686
2451 0a Highly sensitive and specific protein detection via proximity ligation in capillary westerns
338 a print2 rdacarrier
520 a Detection and quantification of proteins and their post-translational modifications are crucial todecipher functions of complex networks in cell biology and medicine. Affinity-based protein analyseshave an important role in proteomic research, and western blotting is one of the most-widely usedtechniques. The NanoPro 1000 system developed by ProteinSimple is an instrument that can resolveand identify proteins and their isoforms through capillary isoelectric focusing with antibody baseddetection directly in the capillaries, in analogy to conventional western blotting. The in situ proximityligation assay (PLA) serves to detect the location of proteins using pairs of oligonucleotide-conjugatedantibodies that mediate formation of rolling circle amplification (RCA) products upon recognition ofthe target protein, or single or dual primary antibodies binding the target. PLA can offer improvedspecificity of detection, and increased sensitivity via, RCA initiated by oligonucleotides attached tothe antibody pairs. Here we have used in situ PLA to detect single or pairs of primary antibodiesbinding proteins separated in the NanoPro 1000 system for highly sensitive and specific detection ofproteins and their isoforms. We achieved at least 10-fold improved detection limits for Erk protein inHDMEC cell lysates, compared with the standard NanoPro 1000 assays, and we demonstrated greatlyenhanced specificity of detection by combining pairs of antibodies, each of which exhibited crossreactivesignals when used on their own.
650 7a NATURVETENSKAPx Biologix Biokemi och molekylärbiologi0 (SwePub)106022 hsv//swe
650 7a NATURAL SCIENCESx Biological Sciencesx Biochemistry and Molecular Biology0 (SwePub)106022 hsv//eng
7001 a Padhan, Narendrau Uppsala universitet,Institutionen för immunologi, genetik och patologi4 aut
7001 a Boge, Annegret4 aut
7001 a Scrivener, Elaine4 aut
7001 a Zieba, Agata4 aut
7001 a Gullberg, Mats4 aut
7001 a Claesson-Welsh, Lenau Uppsala universitet,Institutionen för immunologi, genetik och patologi4 aut
7001 a Kamali-Moghaddam, Masoodu Uppsala universitet,Institutionen för immunologi, genetik och patologi4 aut
7001 a Landegren, Ulfu Uppsala universitet,Institutionen för immunologi, genetik och patologi4 aut
7102 a Uppsala universitetb Institutionen för immunologi, genetik och patologi4 org
8564 8u https://urn.kb.se/resolve?urn=urn:nbn:se:uu:diva-236588

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