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Left top panel: Cartoon representation of E. coli expression and the corresponding 10% Coomassie-stained SDSPAGE gel showing that the purified protein sample contains ATAD2B along with the GroEL contaminant. Left middle panel: Size-exclusion chromatography (Superdex 200 Increase 10/300 GL) profile of the purified sample displaying peaks corresponding to ATAD2B (at 9.01 mLs) and GroEL (at 14.66 mLs). Left bottom panel: Cryo-EM micrograph confirms the presence of heptameric GroEL contaminant in the purified sample. Right top panel: Cartoon representation of the <t>Sf9</t> expression system and the corresponding 10% Coomassiestained SDS-PAGE gel showing that the purified protein sample contains more than 90% pure ATAD2B. Right middle panel: Size-exclusion chromatography (Superdex 200 Increase 10/300 GL) profile of the purified sample showing peaks corresponding to ATAD2B (at 9.01 mLs) and the cleaved GST tag (at 15.89 mLs). Right bottom panel: Cryo-EM micrograph confirms the presence of hexameric ATAD2B in the purified sample.
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Left top panel: Cartoon representation of E. coli expression and the corresponding 10% Coomassie-stained SDSPAGE gel showing that the purified protein sample contains ATAD2B along with the GroEL contaminant. Left middle panel: Size-exclusion chromatography (Superdex 200 Increase 10/300 GL) profile of the purified sample displaying peaks corresponding to ATAD2B (at 9.01 mLs) and GroEL (at 14.66 mLs). Left bottom panel: Cryo-EM micrograph confirms the presence of heptameric GroEL contaminant in the purified sample. Right top panel: Cartoon representation of the <t>Sf9</t> expression system and the corresponding 10% Coomassiestained SDS-PAGE gel showing that the purified protein sample contains more than 90% pure ATAD2B. Right middle panel: Size-exclusion chromatography (Superdex 200 Increase 10/300 GL) profile of the purified sample showing peaks corresponding to ATAD2B (at 9.01 mLs) and the cleaved GST tag (at 15.89 mLs). Right bottom panel: Cryo-EM micrograph confirms the presence of hexameric ATAD2B in the purified sample.
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Left top panel: Cartoon representation of E. coli expression and the corresponding 10% Coomassie-stained SDSPAGE gel showing that the purified protein sample contains ATAD2B along with the GroEL contaminant. Left middle panel: Size-exclusion chromatography (Superdex 200 Increase 10/300 GL) profile of the purified sample displaying peaks corresponding to ATAD2B (at 9.01 mLs) and GroEL (at 14.66 mLs). Left bottom panel: Cryo-EM micrograph confirms the presence of heptameric GroEL contaminant in the purified sample. Right top panel: Cartoon representation of the Sf9 expression system and the corresponding 10% Coomassiestained SDS-PAGE gel showing that the purified protein sample contains more than 90% pure ATAD2B. Right middle panel: Size-exclusion chromatography (Superdex 200 Increase 10/300 GL) profile of the purified sample showing peaks corresponding to ATAD2B (at 9.01 mLs) and the cleaved GST tag (at 15.89 mLs). Right bottom panel: Cryo-EM micrograph confirms the presence of hexameric ATAD2B in the purified sample.

Journal: bioRxiv

Article Title: Breaking Barriers: Transitioning from X-ray Crystallography to Cryo-EM for Structural Studies of ATAD2B

doi: 10.1101/2025.10.13.682238

Figure Lengend Snippet: Left top panel: Cartoon representation of E. coli expression and the corresponding 10% Coomassie-stained SDSPAGE gel showing that the purified protein sample contains ATAD2B along with the GroEL contaminant. Left middle panel: Size-exclusion chromatography (Superdex 200 Increase 10/300 GL) profile of the purified sample displaying peaks corresponding to ATAD2B (at 9.01 mLs) and GroEL (at 14.66 mLs). Left bottom panel: Cryo-EM micrograph confirms the presence of heptameric GroEL contaminant in the purified sample. Right top panel: Cartoon representation of the Sf9 expression system and the corresponding 10% Coomassiestained SDS-PAGE gel showing that the purified protein sample contains more than 90% pure ATAD2B. Right middle panel: Size-exclusion chromatography (Superdex 200 Increase 10/300 GL) profile of the purified sample showing peaks corresponding to ATAD2B (at 9.01 mLs) and the cleaved GST tag (at 15.89 mLs). Right bottom panel: Cryo-EM micrograph confirms the presence of hexameric ATAD2B in the purified sample.

Article Snippet: In the literature, cryo-EM structures of ATAD2 homologs Abo1 and Yta7 were successfully determined from protein samples generated with the Sf9 insect cell expression system ( Cho et al ., 2019 , Cho et al ., 2023 , Wang et al ., 2023 ).

Techniques: Expressing, Staining, Purification, Size-exclusion Chromatography, Cryo-EM Sample Prep, SDS Page