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 trans-Golgi network (TGN) is critical for the biogenesis of secretory granules. Here, we describe a protocol to purify the tagged version of chromogra...

Full description

Saved in:
Bibliographic Details
Main Authors: Anup Parchure, Julia von Blume
Format: Article
Language:English
Published: Bio-protocol LLC 2024-10-01
Series:Bio-Protocol
Online Access:https://bio-protocol.org/en/bpdetail?id=5095&type=0
Tags: Add Tag
No Tags, Be the first to tag this record!
_version_ 1850253399814045696
author Anup Parchure
Julia von Blume
author_facet Anup Parchure
Julia von Blume
author_sort Anup Parchure
collection DOAJ
description Chromogranin B and other members of the granin protein family form condensates that recruit clients like proinsulin. The condensation in the lumen of trans-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.
format Article
id doaj-art-906a0d88bf7f403c888f45c45e5fdaf8
institution OA Journals
issn 2331-8325
language English
publishDate 2024-10-01
publisher Bio-protocol LLC
record_format Article
series Bio-Protocol
spelling doaj-art-906a0d88bf7f403c888f45c45e5fdaf82025-08-20T01:57:24ZengBio-protocol LLCBio-Protocol2331-83252024-10-01142010.21769/BioProtoc.5095Chromogranin B Purification for Condensate Formation and Client Partitioning Assays In VitroAnup Parchure0Julia von Blume1Section of Endocrinology, Department of Internal Medicine, Yale School of Medicine, New Haven, CT, USADepartment of Cell Biology, Yale School of Medicine, New Haven, CT, USADepartment of Cell Biology, Yale School of Medicine, New Haven, CT, USAChromogranin B and other members of the granin protein family form condensates that recruit clients like proinsulin. The condensation in the lumen of trans-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.https://bio-protocol.org/en/bpdetail?id=5095&type=0
spellingShingle Anup Parchure
Julia von Blume
Chromogranin B Purification for Condensate Formation and Client Partitioning Assays In Vitro
Bio-Protocol
title Chromogranin B Purification for Condensate Formation and Client Partitioning Assays In Vitro
title_full Chromogranin B Purification for Condensate Formation and Client Partitioning Assays In Vitro
title_fullStr Chromogranin B Purification for Condensate Formation and Client Partitioning Assays In Vitro
title_full_unstemmed Chromogranin B Purification for Condensate Formation and Client Partitioning Assays In Vitro
title_short Chromogranin B Purification for Condensate Formation and Client Partitioning Assays In Vitro
title_sort chromogranin b purification for condensate formation and client partitioning assays in vitro
url https://bio-protocol.org/en/bpdetail?id=5095&type=0
work_keys_str_mv AT anupparchure chromograninbpurificationforcondensateformationandclientpartitioningassaysinvitro
AT juliavonblume chromograninbpurificationforcondensateformationandclientpartitioningassaysinvitro