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b 20 mm tris ph 8 0 400 mm nacl 250 mm imidazole  (Thermo Fisher)
86
Thermo Fisher b 20 mm tris ph 8 0 400 mm nacl 250 mm imidazole
( A ) Probabilities of individual Met transcription factors (TFs) (Met4, Met32, Met31, Met28), GFP, and mCherry to undergo LLPS from a published prediction program (Fuzdrop). ( B ) Coomassie staining and western blots of purified Met4 and Met32 proteins. For each protein, three purifications were performed and individually imaged. ( C ) Protein aggregate and droplet formation observed for Met4-MBP and Met32-mCherry fusion proteins. Met4 and GFP are 5 µM, Met32-mCherry and mCherry are 10 µM in <t>20</t> <t>mM</t> HEPES (2-(4-(2-hydroxyethyl)piperazin-1-yl)ethanesulfonic acid) pH 7.5, 150 mM NaCl buffer. Met4 and GFP are in 20 mM HEPES pH 7.5, 150 mM NaCl or 50 mM NaCl. Met4 is labeled with Alexa 488 (AF488). Scale bar represents 10 µm (same as in D–F). ( D ) Merging of Met32-mCherry droplets. Met32-mCherry is at 30 µM in 20 mM HEPES pH 7.5, 150 mM NaCl. ( E ) Fluorescence recovery after photobleaching (FRAP) data for 20 µM Met32 droplets. The intensity data was collected every 3 s for 270 s and normalized to percent bleaching. Error bars represent the standard deviation of three biological replicates. Inset: Representative images of Met32 FRAP. ( F ) Co-localization of Met32-mCherry droplets (10 µM) with Met4-MBP (5 µM) in 150 mM NaCl. Figure 2—source data 1. Raw Gel Blots. Figure 2—source data 2. Labeled Gel Blots.
B 20 Mm Tris Ph 8 0 400 Mm Nacl 250 Mm Imidazole, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/result/b 20 mm tris ph 8 0 400 mm nacl 250 mm imidazole/product/Thermo Fisher
Average 86 stars, based on 1 article reviews
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b 400 07 003  (Foss Analytical)
86
Foss Analytical b 400 07 003
( A ) Probabilities of individual Met transcription factors (TFs) (Met4, Met32, Met31, Met28), GFP, and mCherry to undergo LLPS from a published prediction program (Fuzdrop). ( B ) Coomassie staining and western blots of purified Met4 and Met32 proteins. For each protein, three purifications were performed and individually imaged. ( C ) Protein aggregate and droplet formation observed for Met4-MBP and Met32-mCherry fusion proteins. Met4 and GFP are 5 µM, Met32-mCherry and mCherry are 10 µM in <t>20</t> <t>mM</t> HEPES (2-(4-(2-hydroxyethyl)piperazin-1-yl)ethanesulfonic acid) pH 7.5, 150 mM NaCl buffer. Met4 and GFP are in 20 mM HEPES pH 7.5, 150 mM NaCl or 50 mM NaCl. Met4 is labeled with Alexa 488 (AF488). Scale bar represents 10 µm (same as in D–F). ( D ) Merging of Met32-mCherry droplets. Met32-mCherry is at 30 µM in 20 mM HEPES pH 7.5, 150 mM NaCl. ( E ) Fluorescence recovery after photobleaching (FRAP) data for 20 µM Met32 droplets. The intensity data was collected every 3 s for 270 s and normalized to percent bleaching. Error bars represent the standard deviation of three biological replicates. Inset: Representative images of Met32 FRAP. ( F ) Co-localization of Met32-mCherry droplets (10 µM) with Met4-MBP (5 µM) in 150 mM NaCl. Figure 2—source data 1. Raw Gel Blots. Figure 2—source data 2. Labeled Gel Blots.
B 400 07 003, supplied by Foss Analytical, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/result/b 400 07 003/product/Foss Analytical
Average 86 stars, based on 1 article reviews
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b 400 07 003 - by Bioz Stars, 2024-10
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400 mhz ultrashield plus b acs 60 spectrometer  (Bruker Corporation)
86
Bruker Corporation 400 mhz ultrashield plus b acs 60 spectrometer
( A ) Probabilities of individual Met transcription factors (TFs) (Met4, Met32, Met31, Met28), GFP, and mCherry to undergo LLPS from a published prediction program (Fuzdrop). ( B ) Coomassie staining and western blots of purified Met4 and Met32 proteins. For each protein, three purifications were performed and individually imaged. ( C ) Protein aggregate and droplet formation observed for Met4-MBP and Met32-mCherry fusion proteins. Met4 and GFP are 5 µM, Met32-mCherry and mCherry are 10 µM in <t>20</t> <t>mM</t> HEPES (2-(4-(2-hydroxyethyl)piperazin-1-yl)ethanesulfonic acid) pH 7.5, 150 mM NaCl buffer. Met4 and GFP are in 20 mM HEPES pH 7.5, 150 mM NaCl or 50 mM NaCl. Met4 is labeled with Alexa 488 (AF488). Scale bar represents 10 µm (same as in D–F). ( D ) Merging of Met32-mCherry droplets. Met32-mCherry is at 30 µM in 20 mM HEPES pH 7.5, 150 mM NaCl. ( E ) Fluorescence recovery after photobleaching (FRAP) data for 20 µM Met32 droplets. The intensity data was collected every 3 s for 270 s and normalized to percent bleaching. Error bars represent the standard deviation of three biological replicates. Inset: Representative images of Met32 FRAP. ( F ) Co-localization of Met32-mCherry droplets (10 µM) with Met4-MBP (5 µM) in 150 mM NaCl. Figure 2—source data 1. Raw Gel Blots. Figure 2—source data 2. Labeled Gel Blots.
400 Mhz Ultrashield Plus B Acs 60 Spectrometer, supplied by Bruker Corporation, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/result/400 mhz ultrashield plus b acs 60 spectrometer/product/Bruker Corporation
Average 86 stars, based on 1 article reviews
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400 mhz ultrashield plus b acs 60 spectrometer - by Bioz Stars, 2024-10
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dfl newdoon foc c b 400 1 25 0 50 1 5 rhd 32 channel recording headstages intan technologies part  (NEWDOON Inc)
86
NEWDOON Inc dfl newdoon foc c b 400 1 25 0 50 1 5 rhd 32 channel recording headstages intan technologies part
( A ) Probabilities of individual Met transcription factors (TFs) (Met4, Met32, Met31, Met28), GFP, and mCherry to undergo LLPS from a published prediction program (Fuzdrop). ( B ) Coomassie staining and western blots of purified Met4 and Met32 proteins. For each protein, three purifications were performed and individually imaged. ( C ) Protein aggregate and droplet formation observed for Met4-MBP and Met32-mCherry fusion proteins. Met4 and GFP are 5 µM, Met32-mCherry and mCherry are 10 µM in <t>20</t> <t>mM</t> HEPES (2-(4-(2-hydroxyethyl)piperazin-1-yl)ethanesulfonic acid) pH 7.5, 150 mM NaCl buffer. Met4 and GFP are in 20 mM HEPES pH 7.5, 150 mM NaCl or 50 mM NaCl. Met4 is labeled with Alexa 488 (AF488). Scale bar represents 10 µm (same as in D–F). ( D ) Merging of Met32-mCherry droplets. Met32-mCherry is at 30 µM in 20 mM HEPES pH 7.5, 150 mM NaCl. ( E ) Fluorescence recovery after photobleaching (FRAP) data for 20 µM Met32 droplets. The intensity data was collected every 3 s for 270 s and normalized to percent bleaching. Error bars represent the standard deviation of three biological replicates. Inset: Representative images of Met32 FRAP. ( F ) Co-localization of Met32-mCherry droplets (10 µM) with Met4-MBP (5 µM) in 150 mM NaCl. Figure 2—source data 1. Raw Gel Blots. Figure 2—source data 2. Labeled Gel Blots.
Dfl Newdoon Foc C B 400 1 25 0 50 1 5 Rhd 32 Channel Recording Headstages Intan Technologies Part, supplied by NEWDOON Inc, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/result/dfl newdoon foc c b 400 1 25 0 50 1 5 rhd 32 channel recording headstages intan technologies part/product/NEWDOON Inc
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dfl newdoon foc c b 400 1 25 0 50 1 5 rhd 32 channel recording headstages intan technologies part - by Bioz Stars, 2024-10
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ccd camera roper scientific spec 10 400 b ln snu  (Roper Scientific Inc)
86
Roper Scientific Inc ccd camera roper scientific spec 10 400 b ln snu
( A ) Probabilities of individual Met transcription factors (TFs) (Met4, Met32, Met31, Met28), GFP, and mCherry to undergo LLPS from a published prediction program (Fuzdrop). ( B ) Coomassie staining and western blots of purified Met4 and Met32 proteins. For each protein, three purifications were performed and individually imaged. ( C ) Protein aggregate and droplet formation observed for Met4-MBP and Met32-mCherry fusion proteins. Met4 and GFP are 5 µM, Met32-mCherry and mCherry are 10 µM in <t>20</t> <t>mM</t> HEPES (2-(4-(2-hydroxyethyl)piperazin-1-yl)ethanesulfonic acid) pH 7.5, 150 mM NaCl buffer. Met4 and GFP are in 20 mM HEPES pH 7.5, 150 mM NaCl or 50 mM NaCl. Met4 is labeled with Alexa 488 (AF488). Scale bar represents 10 µm (same as in D–F). ( D ) Merging of Met32-mCherry droplets. Met32-mCherry is at 30 µM in 20 mM HEPES pH 7.5, 150 mM NaCl. ( E ) Fluorescence recovery after photobleaching (FRAP) data for 20 µM Met32 droplets. The intensity data was collected every 3 s for 270 s and normalized to percent bleaching. Error bars represent the standard deviation of three biological replicates. Inset: Representative images of Met32 FRAP. ( F ) Co-localization of Met32-mCherry droplets (10 µM) with Met4-MBP (5 µM) in 150 mM NaCl. Figure 2—source data 1. Raw Gel Blots. Figure 2—source data 2. Labeled Gel Blots.
Ccd Camera Roper Scientific Spec 10 400 B Ln Snu, supplied by Roper Scientific Inc, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/result/ccd camera roper scientific spec 10 400 b ln snu/product/Roper Scientific Inc
Average 86 stars, based on 1 article reviews
Price from $9.99 to $1999.99
ccd camera roper scientific spec 10 400 b ln snu - by Bioz Stars, 2024-10
86/100 stars
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400 methanol hexane water ascorbic acid ic 50 b  (GraphPad Software Inc)
86
GraphPad Software Inc 400 methanol hexane water ascorbic acid ic 50 b
( A ) Probabilities of individual Met transcription factors (TFs) (Met4, Met32, Met31, Met28), GFP, and mCherry to undergo LLPS from a published prediction program (Fuzdrop). ( B ) Coomassie staining and western blots of purified Met4 and Met32 proteins. For each protein, three purifications were performed and individually imaged. ( C ) Protein aggregate and droplet formation observed for Met4-MBP and Met32-mCherry fusion proteins. Met4 and GFP are 5 µM, Met32-mCherry and mCherry are 10 µM in <t>20</t> <t>mM</t> HEPES (2-(4-(2-hydroxyethyl)piperazin-1-yl)ethanesulfonic acid) pH 7.5, 150 mM NaCl buffer. Met4 and GFP are in 20 mM HEPES pH 7.5, 150 mM NaCl or 50 mM NaCl. Met4 is labeled with Alexa 488 (AF488). Scale bar represents 10 µm (same as in D–F). ( D ) Merging of Met32-mCherry droplets. Met32-mCherry is at 30 µM in 20 mM HEPES pH 7.5, 150 mM NaCl. ( E ) Fluorescence recovery after photobleaching (FRAP) data for 20 µM Met32 droplets. The intensity data was collected every 3 s for 270 s and normalized to percent bleaching. Error bars represent the standard deviation of three biological replicates. Inset: Representative images of Met32 FRAP. ( F ) Co-localization of Met32-mCherry droplets (10 µM) with Met4-MBP (5 µM) in 150 mM NaCl. Figure 2—source data 1. Raw Gel Blots. Figure 2—source data 2. Labeled Gel Blots.
400 Methanol Hexane Water Ascorbic Acid Ic 50 B, supplied by GraphPad Software Inc, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/result/400 methanol hexane water ascorbic acid ic 50 b/product/GraphPad Software Inc
Average 86 stars, based on 1 article reviews
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400 methanol hexane water ascorbic acid ic 50 b - by Bioz Stars, 2024-10
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granzyme b biolegend 372210 qa16a02 1 400  (Revvity Signals)
86
Revvity Signals granzyme b biolegend 372210 qa16a02 1 400
( A ) Probabilities of individual Met transcription factors (TFs) (Met4, Met32, Met31, Met28), GFP, and mCherry to undergo LLPS from a published prediction program (Fuzdrop). ( B ) Coomassie staining and western blots of purified Met4 and Met32 proteins. For each protein, three purifications were performed and individually imaged. ( C ) Protein aggregate and droplet formation observed for Met4-MBP and Met32-mCherry fusion proteins. Met4 and GFP are 5 µM, Met32-mCherry and mCherry are 10 µM in <t>20</t> <t>mM</t> HEPES (2-(4-(2-hydroxyethyl)piperazin-1-yl)ethanesulfonic acid) pH 7.5, 150 mM NaCl buffer. Met4 and GFP are in 20 mM HEPES pH 7.5, 150 mM NaCl or 50 mM NaCl. Met4 is labeled with Alexa 488 (AF488). Scale bar represents 10 µm (same as in D–F). ( D ) Merging of Met32-mCherry droplets. Met32-mCherry is at 30 µM in 20 mM HEPES pH 7.5, 150 mM NaCl. ( E ) Fluorescence recovery after photobleaching (FRAP) data for 20 µM Met32 droplets. The intensity data was collected every 3 s for 270 s and normalized to percent bleaching. Error bars represent the standard deviation of three biological replicates. Inset: Representative images of Met32 FRAP. ( F ) Co-localization of Met32-mCherry droplets (10 µM) with Met4-MBP (5 µM) in 150 mM NaCl. Figure 2—source data 1. Raw Gel Blots. Figure 2—source data 2. Labeled Gel Blots.
Granzyme B Biolegend 372210 Qa16a02 1 400, supplied by Revvity Signals, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/result/granzyme b biolegend 372210 qa16a02 1 400/product/Revvity Signals
Average 86 stars, based on 1 article reviews
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granzyme b biolegend 372210 qa16a02 1 400 - by Bioz Stars, 2024-10
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y 0 200 400 600 cal np b fr eq ue nc y  (Lonza)
86
Lonza y 0 200 400 600 cal np b fr eq ue nc y
( A ) Probabilities of individual Met transcription factors (TFs) (Met4, Met32, Met31, Met28), GFP, and mCherry to undergo LLPS from a published prediction program (Fuzdrop). ( B ) Coomassie staining and western blots of purified Met4 and Met32 proteins. For each protein, three purifications were performed and individually imaged. ( C ) Protein aggregate and droplet formation observed for Met4-MBP and Met32-mCherry fusion proteins. Met4 and GFP are 5 µM, Met32-mCherry and mCherry are 10 µM in <t>20</t> <t>mM</t> HEPES (2-(4-(2-hydroxyethyl)piperazin-1-yl)ethanesulfonic acid) pH 7.5, 150 mM NaCl buffer. Met4 and GFP are in 20 mM HEPES pH 7.5, 150 mM NaCl or 50 mM NaCl. Met4 is labeled with Alexa 488 (AF488). Scale bar represents 10 µm (same as in D–F). ( D ) Merging of Met32-mCherry droplets. Met32-mCherry is at 30 µM in 20 mM HEPES pH 7.5, 150 mM NaCl. ( E ) Fluorescence recovery after photobleaching (FRAP) data for 20 µM Met32 droplets. The intensity data was collected every 3 s for 270 s and normalized to percent bleaching. Error bars represent the standard deviation of three biological replicates. Inset: Representative images of Met32 FRAP. ( F ) Co-localization of Met32-mCherry droplets (10 µM) with Met4-MBP (5 µM) in 150 mM NaCl. Figure 2—source data 1. Raw Gel Blots. Figure 2—source data 2. Labeled Gel Blots.
Y 0 200 400 600 Cal Np B Fr Eq Ue Nc Y, supplied by Lonza, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/result/y 0 200 400 600 cal np b fr eq ue nc y/product/Lonza
Average 86 stars, based on 1 article reviews
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ti o n i n n g l b dieldrin β hch 1 cl cldoh a1 a2 cldoh 0 400 800 1200 c o n c e n tr  (Genoscope)
86
Genoscope ti o n i n n g l b dieldrin β hch 1 cl cldoh a1 a2 cldoh 0 400 800 1200 c o n c e n tr
( A ) Probabilities of individual Met transcription factors (TFs) (Met4, Met32, Met31, Met28), GFP, and mCherry to undergo LLPS from a published prediction program (Fuzdrop). ( B ) Coomassie staining and western blots of purified Met4 and Met32 proteins. For each protein, three purifications were performed and individually imaged. ( C ) Protein aggregate and droplet formation observed for Met4-MBP and Met32-mCherry fusion proteins. Met4 and GFP are 5 µM, Met32-mCherry and mCherry are 10 µM in <t>20</t> <t>mM</t> HEPES (2-(4-(2-hydroxyethyl)piperazin-1-yl)ethanesulfonic acid) pH 7.5, 150 mM NaCl buffer. Met4 and GFP are in 20 mM HEPES pH 7.5, 150 mM NaCl or 50 mM NaCl. Met4 is labeled with Alexa 488 (AF488). Scale bar represents 10 µm (same as in D–F). ( D ) Merging of Met32-mCherry droplets. Met32-mCherry is at 30 µM in 20 mM HEPES pH 7.5, 150 mM NaCl. ( E ) Fluorescence recovery after photobleaching (FRAP) data for 20 µM Met32 droplets. The intensity data was collected every 3 s for 270 s and normalized to percent bleaching. Error bars represent the standard deviation of three biological replicates. Inset: Representative images of Met32 FRAP. ( F ) Co-localization of Met32-mCherry droplets (10 µM) with Met4-MBP (5 µM) in 150 mM NaCl. Figure 2—source data 1. Raw Gel Blots. Figure 2—source data 2. Labeled Gel Blots.
Ti O N I N N G L B Dieldrin β Hch 1 Cl Cldoh A1 A2 Cldoh 0 400 800 1200 C O N C E N Tr, supplied by Genoscope, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/result/ti o n i n n g l b dieldrin β hch 1 cl cldoh a1 a2 cldoh 0 400 800 1200 c o n c e n tr/product/Genoscope
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ti o n i n n g l b dieldrin β hch 1 cl cldoh a1 a2 cldoh 0 400 800 1200 c o n c e n tr - by Bioz Stars, 2024-10
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Image Search Results


( A ) Probabilities of individual Met transcription factors (TFs) (Met4, Met32, Met31, Met28), GFP, and mCherry to undergo LLPS from a published prediction program (Fuzdrop). ( B ) Coomassie staining and western blots of purified Met4 and Met32 proteins. For each protein, three purifications were performed and individually imaged. ( C ) Protein aggregate and droplet formation observed for Met4-MBP and Met32-mCherry fusion proteins. Met4 and GFP are 5 µM, Met32-mCherry and mCherry are 10 µM in 20 mM HEPES (2-(4-(2-hydroxyethyl)piperazin-1-yl)ethanesulfonic acid) pH 7.5, 150 mM NaCl buffer. Met4 and GFP are in 20 mM HEPES pH 7.5, 150 mM NaCl or 50 mM NaCl. Met4 is labeled with Alexa 488 (AF488). Scale bar represents 10 µm (same as in D–F). ( D ) Merging of Met32-mCherry droplets. Met32-mCherry is at 30 µM in 20 mM HEPES pH 7.5, 150 mM NaCl. ( E ) Fluorescence recovery after photobleaching (FRAP) data for 20 µM Met32 droplets. The intensity data was collected every 3 s for 270 s and normalized to percent bleaching. Error bars represent the standard deviation of three biological replicates. Inset: Representative images of Met32 FRAP. ( F ) Co-localization of Met32-mCherry droplets (10 µM) with Met4-MBP (5 µM) in 150 mM NaCl. Figure 2—source data 1. Raw Gel Blots. Figure 2—source data 2. Labeled Gel Blots.

Journal: eLife

Article Title: Transcription factor condensates, 3D clustering, and gene expression enhancement of the MET regulon

doi: 10.7554/eLife.96028

Figure Lengend Snippet: ( A ) Probabilities of individual Met transcription factors (TFs) (Met4, Met32, Met31, Met28), GFP, and mCherry to undergo LLPS from a published prediction program (Fuzdrop). ( B ) Coomassie staining and western blots of purified Met4 and Met32 proteins. For each protein, three purifications were performed and individually imaged. ( C ) Protein aggregate and droplet formation observed for Met4-MBP and Met32-mCherry fusion proteins. Met4 and GFP are 5 µM, Met32-mCherry and mCherry are 10 µM in 20 mM HEPES (2-(4-(2-hydroxyethyl)piperazin-1-yl)ethanesulfonic acid) pH 7.5, 150 mM NaCl buffer. Met4 and GFP are in 20 mM HEPES pH 7.5, 150 mM NaCl or 50 mM NaCl. Met4 is labeled with Alexa 488 (AF488). Scale bar represents 10 µm (same as in D–F). ( D ) Merging of Met32-mCherry droplets. Met32-mCherry is at 30 µM in 20 mM HEPES pH 7.5, 150 mM NaCl. ( E ) Fluorescence recovery after photobleaching (FRAP) data for 20 µM Met32 droplets. The intensity data was collected every 3 s for 270 s and normalized to percent bleaching. Error bars represent the standard deviation of three biological replicates. Inset: Representative images of Met32 FRAP. ( F ) Co-localization of Met32-mCherry droplets (10 µM) with Met4-MBP (5 µM) in 150 mM NaCl. Figure 2—source data 1. Raw Gel Blots. Figure 2—source data 2. Labeled Gel Blots.

Article Snippet: Met32-mCherry was purified using Ni-NTA chromatography (Thermo Scientific HisPur resin #88222) with proteins buffers A (20 mM Tris pH 8.0, 400 mM NaCl, 5 mM imidazole) and B (20 mM Tris pH 8.0, 400 mM NaCl, 250 mM imidazole) with a stepwise gradient of buffer B.

Techniques: Staining, Western Blot, Purification, Labeling, Fluorescence, Standard Deviation

( A ) Predicted protein structures of Met4 (Alphafold: AF-A0A1L4AA63-F1) and Met32 (AF-A0A0L8VTL7-F1). ( B ) PONDR protein disorder plot of Met32 (same analysis for Met4 is shown in ). ( C ) Met32-mCherry condensate formation at various concentrations. Purified Met32-mCherry proteins were titrated in 20 mM HEPES pH 7.5, 150 mM NaCl. Scale bar represents 10 µm. ( D ) Fluorescence recovery after photobleaching (FRAP) curve for Met4/Met32 co-localized droplets. 5 µM Met4-MBP and 20 µM Met32-mCherry were mixed. Intensity data collected every 3 s for 270 s.

Journal: eLife

Article Title: Transcription factor condensates, 3D clustering, and gene expression enhancement of the MET regulon

doi: 10.7554/eLife.96028

Figure Lengend Snippet: ( A ) Predicted protein structures of Met4 (Alphafold: AF-A0A1L4AA63-F1) and Met32 (AF-A0A0L8VTL7-F1). ( B ) PONDR protein disorder plot of Met32 (same analysis for Met4 is shown in ). ( C ) Met32-mCherry condensate formation at various concentrations. Purified Met32-mCherry proteins were titrated in 20 mM HEPES pH 7.5, 150 mM NaCl. Scale bar represents 10 µm. ( D ) Fluorescence recovery after photobleaching (FRAP) curve for Met4/Met32 co-localized droplets. 5 µM Met4-MBP and 20 µM Met32-mCherry were mixed. Intensity data collected every 3 s for 270 s.

Article Snippet: Met32-mCherry was purified using Ni-NTA chromatography (Thermo Scientific HisPur resin #88222) with proteins buffers A (20 mM Tris pH 8.0, 400 mM NaCl, 5 mM imidazole) and B (20 mM Tris pH 8.0, 400 mM NaCl, 250 mM imidazole) with a stepwise gradient of buffer B.

Techniques: Purification, Fluorescence