Re: [ccp4bb] Ecoli protein staining with AntiHis?

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From: Peter Adrian Meyer pam52 {- at -} CORNELL {- dot -} EDU
Date: 2007-04-20

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Subject: Re: Ecoli protein staining with AntiHis?
From: Artem Evdokimov artem {- at -} XTALS {- dot -} ORG
Date: 2007-04-20

Depends on which antibody you're using. Different manufacturers tend to
light up different stuff. We very rarely see just one band - typically it's
a bunch of weak bands as well as smears.

How strong is the band? Can you cut it out of the gel and do a tryptic
digest and MS identification? If the band is strong, consider the
possibility that you've somehow contaminated your strain with some other
expressor...

Artem

-----Original Message-----
From: CCP4 bulletin board [mailto:CCP4BB@JISCMAIL.AC.UK] On Behalf Of Raji
Edayathumangalam
Sent: Friday, April 20, 2007 4:36 PM
To: CCP4BB@JISCMAIL.AC.UK
Subject: [ccp4bb] Ecoli protein staining with AntiHis?

Hi Everyone,

I see a band that lights up in my anti-His Western blots.

While I investigate what else the band might be (truncated protein etc.),
does anyone know whether
there is an E. coli protein that migrates ~ 30-40kDa (SDS-PAGE), which binds
to anti-His antibodies?

Thanks.
Raji

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CCP4bb <-- 2007 <-- April 2007 <-- 20 April 2007

Previous message:
Subject: Re: How to run SIGMAA twice in a row? (fwd)
From: Peter Adrian Meyer pam52 {- at -} CORNELL {- dot -} EDU
Date: 2007-04-20