Re: [ccp4bb] Manual His-Tag Purification

2017-02-02 Thread Sundaram
Thanks everyone! Thanks Prof. David Blum. I will do batch load and check. Yours Sincerely, Sundaram.S On Wed, Feb 1, 2017 at 9:06 PM, David Blum wrote: > Hi Sundaram, > > The binding capacity of this column is 40 mg/mL of resin so a 5mL column > will hold a maximum of 200 mg of protein. If y

Re: [ccp4bb] Manual His-Tag Purification

2017-02-01 Thread David Blum
Hi Sundaram, The binding capacity of this column is 40 mg/mL of resin so a 5mL column will hold a maximum of 200 mg of protein. If you run your cleared lysate on a gel you may be able to estimate how much protein there is. Our facility purifies a range of different proteins for investigators and

Re: [ccp4bb] Manual His-Tag Purification

2017-02-01 Thread Tim Gruene
Dear Sundaram S, during my PhD I used 4-7.5ml resin per l of culture, but I also had a large yield of 60-100mg protein per litre. Try to use as little as possible - at first trial check both flow-through and retentate by SDS-PAGE. (see. p. 40 and 54 of my thesis). My constructs would also bind

Re: [ccp4bb] Manual His-Tag Purification

2017-02-01 Thread Horrell, Sam
in board on behalf of Sundaram Sent: 01 February 2017 09:24:09 To: CCP4BB@JISCMAIL.AC.UK Subject: [ccp4bb] Manual His-Tag Purification Hello , It's an off topic question. I'm planning to do manual purification a 6 his tagged protein of size around 20kDa from 1L E.coli culture using C

[ccp4bb] Manual His-Tag Purification

2017-02-01 Thread Sundaram
Hello , It's an off topic question. I'm planning to do manual purification a 6 his tagged protein of size around 20kDa from 1L E.coli culture using COHISC-RO Roche cOmpleteā„¢ His-Tag Purification Column. Can I get some advice regarding the lysate loading volume and retention time. This is the fi