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838 bytes added ,  11:26, 10 February 2008
New page: This protocol is an NKI local adaptation based on the article by Grimes and co-workers[http://www.ncbi.nlm.nih.gov/pubmed/17098187?ordinalpos=1&itool=EntrezSystem2.PEntrez.Pubmed.Pubmed_Re...
This protocol is an NKI local adaptation based on the article by Grimes and co-workers[http://www.ncbi.nlm.nih.gov/pubmed/17098187?ordinalpos=1&itool=EntrezSystem2.PEntrez.Pubmed.Pubmed_ResultsPanel.Pubmed_RVDocSum]

*1mg/ml protein
*50mM HEPES (pH7.5)
*250mM NaCl
*'''Do NOT use Tris'''

Add per 1ml protein:
*20μl 1M dimethylamine-borane complex (ABC) – freshly prepared
*40μl 1M formaldehyde (from 37% stock)

Mix gently and incubate for 2hrs at 4°C

Add again:

*20μl 1M dimethylamine-borane complex
*40μl 1M formaldehyde

Continue incubation for 2hrs at 4°C

Then add

*10μl 1M dimethylamine-borane complex (ABC)

Incubate O/N at 4°C

Remove precipitated protein (up to 50%) by centrifugation

Concentrate and put protein on Gel Filtration column in a Tris conatining buffer, eg:

*20mM Tris (pH7.5)
*200mM NaCl
*1mM DTT
84

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