Centrifugal Protein Concentration and Buffer Exchange - Revision history

Mjeppesen at 20:30, 21 July 2022

2022-07-21T20:30:10Z

← Older revision		Revision as of 20:30, 21 July 2022
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	# Keep the filter wet with 20% ethanol when it is stored.		# Keep the filter wet with 20% ethanol when it is stored.

	~~[[category:Protocols]]~~
	[[category:Protein_Preparation]]		[[category:Protein_Preparation]]

Mjeppesen: LearningSorting

2021-09-11T04:36:28Z

LearningSorting

← Older revision		Revision as of 04:36, 11 September 2021
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	# Keep the filter wet with 20% ethanol when it is stored.		# Keep the filter wet with 20% ethanol when it is stored.

	[[category:~~Protocols\|~~Protocols]]		[[category:Protocols]]
	[[category:Protein_Preparation]]		[[category:Protein_Preparation]]

Mjeppesen: Added to Category

2021-09-11T03:13:34Z

Added to Category

← Older revision		Revision as of 03:13, 11 September 2021
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	## Be sure not to over concentrate and aggregate your protein.		## Be sure not to over concentrate and aggregate your protein.
	# Keep the filter wet with 20% ethanol when it is stored.		# Keep the filter wet with 20% ethanol when it is stored.

			[[category:Protocols\|Protocols]]
			[[category:Protein_Preparation]]

Jcatazaro: Created page with "Centrifugal Protein Concentration/Buffer Exchange # Obtain a centrifugal filter (Amicon or Centricon) with a molecular weight cutoff at least half of your protein’s weight...."

2017-03-14T21:00:55Z

Created page with "Centrifugal Protein Concentration/Buffer Exchange # Obtain a centrifugal filter (Amicon or Centricon) with a molecular weight cutoff at least half of your protein’s weight...."

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Centrifugal Protein Concentration/Buffer Exchange

# Obtain a centrifugal filter (Amicon or Centricon) with a molecular weight cutoff at least half of your protein’s weight.
# If the filter is new, run ~5 mL of nanopure water through before adding your protein mixture.
## Make sure to balance the centrifuge with another centrifugal filter. I typically use an old one. Regular falcon tubes will unbalance the centrifuge even though they are the same weight.
# Add your protein mixture up to the recommended line.
# Spin at 4,000 rpm for ~40 minutes.
# Pour out the filtrate. The supernatant contains your protein.
# Add the rest of your protein mixture or add in the new buffer.
# Continue to add and spin down the new buffer until it is sufficiently exchanged and the protein is concentrated.
## ~50 mL works well
## Be sure not to over concentrate and aggregate your protein.
# Keep the filter wet with 20% ethanol when it is stored.