Chromogranin B Purification for Condensate Formation and Client Partitioning Assays In Vitro
Chromogranin B and other members of the granin protein family form condensates that recruit clients like proinsulin. The condensation in the lumen of -Golgi network (TGN) is critical for the biogenesis of secretory granules. Here, we describe a protocol to purify the tagged version of chromogranin B...
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Veröffentlicht in: | Bio-protocol 2024-10, Vol.14 (20), p.e5095 |
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Format: | Artikel |
Sprache: | eng |
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Zusammenfassung: | Chromogranin B and other members of the granin protein family form condensates that recruit clients like proinsulin. The condensation in the lumen of
-Golgi network (TGN) is critical for the biogenesis of secretory granules. Here, we describe a protocol to purify the tagged version of chromogranin B close to its native form at the TGN, which can then be utilized for microscopy-based assays to monitor condensate formation in vitro and client partitioning depending on the material properties of chromogranin B assemblies. Key features • First instance of purification of full-length and tagged version of members of the chromogranin family of proteins. • Allows purification of proteins with post-translational modifications that are acquired en route in the secretory pathway, thus closely resembling their native form at the TGN. |
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ISSN: | 2331-8325 2331-8325 |
DOI: | 10.21769/BioProtoc.5095 |