Re: [ccp4bb] Off-topic: Attach His-tagged Protein to Coverslip

2017-04-05 Thread Tristan Croll 
One possibility: surface-immobilised forms of the peptide HGGHHG bind strongly 
to His tags (coordinating around zinc rather than nickel). See 
https://www.ncbi.nlm.nih.gov/m/pubmed/15050643/. If you want true permanence, 
you could apply the trick used on BiaCore chips. They use a NTA-functional 
carboxymethyldextran surface. His-tagged proteins are captured as usual, then 
rinsed and made permanent by incubating with some water-soluble carbodiimide to 
link the carboxymethyl groups to surface lysines - the advantage being that all 
the proteins are in essentially the same orientation. Not sure if anyone sells 
coverslip equivalents.

Best regards,

Tristan

 
 
Tristan Croll
Research Fellow
Cambridge Institute for Medical Research
University of Cambridge CB2 0XY
 

 

> On 5 Apr 2017, at 04:19, Keller, Jacob  wrote:
> 
> Does anyone have a simple way to attach purified his-tagged protein solidly 
> to a coverslip?
>  
> Thanks,
>  
> Jacob Keller
>  
> ***
> Jacob Pearson Keller, PhD
> Research Scientist
> HHMI Janelia Research Campus / Looger lab
> Phone: (571)209-4000 x3159
> Email: kell...@janelia.hhmi.org
> ***
>

[ccp4bb] Spam : Re: [ccp4bb] Off-topic: Attach His-tagged Protein to Coverslip

2017-04-04 Thread Debasish Kumar Ghosh 
Assuming the protein is in solution, drop cast 2ul of solution onto a glass 
surface and spread evenly with a sharp needle. Air dry in dust free chamber and 
blow the surface with gentle stream of nitrogen gas. Proteins stick to surface 
quite well to be analyzed for various microscopy (AFM, SEM etc.). If the 
purpose is not to do any surface probe microscopy, coating the glass surface 
with Sigmacoat helps in better binding. Poly-lysine coated glass cover slips 
can be alternative but mostly not preferred for small proteins or peptides.

Hope that helps.

Best!!


Debasish

CSIR- Senior Research Fellow (PhD Scholar)
C/o: Dr. Akash Ranjan
Computational and Functional Genomics Group
Centre for DNA Fingerprinting and Diagnostics
Hyderabad, INDIA

Email(s): dkgh...@cdfd.org.in, dgho...@gmail.com
Telephone: 0091-9088334375 (M), 0091-40-24749396 (Lab)
Lab URL: http://www.cdfd.org.in/labpages/computational_functional_genomics.html



- Original Message -
From: Jacob Keller <kell...@janelia.hhmi.org>
To: CCP4BB@JISCMAIL.AC.UK
Sent: Wed, 05 Apr 2017 08:49:40 +0530 (IST)
Subject: [ccp4bb] Off-topic: Attach His-tagged Protein to Coverslip

Does anyone have a simple way to attach purified his-tagged protein solidly to 
a coverslip?

Thanks,

Jacob Keller

***
Jacob Pearson Keller, PhD
Research Scientist
HHMI Janelia Research Campus / Looger lab
Phone: (571)209-4000 x3159
Email: kell...@janelia.hhmi.org<mailto:kell...@janelia.hhmi.org>
***

[ccp4bb] Off-topic: Attach His-tagged Protein to Coverslip

2017-04-04 Thread Keller, Jacob 
Does anyone have a simple way to attach purified his-tagged protein solidly to 
a coverslip?

Thanks,

Jacob Keller

***
Jacob Pearson Keller, PhD
Research Scientist
HHMI Janelia Research Campus / Looger lab
Phone: (571)209-4000 x3159
Email: kell...@janelia.hhmi.org
***