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protein DNA crosslinked complex purification

protein DNA crosslinked complex purification - Protocols and Methods Forum

protein DNA crosslinked complex purification - Post Any Protocol, Method, Technique, Procedure or Tips / Troubleshooting for any Molecular Biology Technique.


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  #1  
Old 10-19-2007, 05:35 AM
qiang fu
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Default protein DNA crosslinked complex purification



Hi, Guys:

I am recently doing some UV crosslink between the protein and DNA.(Histone protein and DNA)
Due to our small amount sample about 50 ul and low percentage of successful crosslink. Is there any method to isolate the crosslinked sample out from the DNA with no protein crosslinked.

Your suggestions will be great helpful.

Thanks a lot!

Qiang,

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  #2  
Old 10-19-2007, 07:08 PM
kaj.stenberg@helsinki.fi.invalid
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Default protein DNA crosslinked complex purification

qiang fu <[Only registered users see links. ]> wrote:

Could you run an EMSA on a preparative scale?
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  #3  
Old 10-22-2007, 05:03 PM
Yuval Ebenstein
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Default Re: protein DNA crosslinked complex purification

If the DNA is Short enough to see a gel shift, do a gel extraction.

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Yuval Ebenstein
Department of Chemistry and Biochemistry, UCLA
Young Hall-2002, 607 Charles E. Young Drive East
Los Angeles, CA 90095-1569
Ph: (310) 794-6685; Fax: (310) 267-4672

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Old 10-22-2007, 11:36 PM
Zhonglin Chai
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Default Re: protein DNA crosslinked complex purification

how about using formaldehyde as used in ChIP assay? directly add
formaldehyde to cultured cells at final 1%, and incubate for 10 min.

Dr Zhonglin Chai, PhD
Head
Cell Proliferation & Fibrosis In Diabetic Complications Laboratory
Diabetes & Metabolism Division
Baker Heart Research Institute
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Tel: +61 3 8532 1231
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Ebenstein
Sent: Tuesday, 23 October 2007 3:04 AM
To: [Only registered users see links. ]lid; [Only registered users see links. ]
Subject: Re: Re: protein DNA crosslinked complex purification

If the DNA is Short enough to see a gel shift, do a gel extraction.

[Only registered users see links. ]lid wrote:
DNA.(Histone protein and DNA)
successful crosslink. Is there any method to isolate the crosslinked
sample out from the DNA with no protein crosslinked.
-------------------------------------------

Yuval Ebenstein
Department of Chemistry and Biochemistry, UCLA Young Hall-2002, 607
Charles E. Young Drive East Los Angeles, CA 90095-1569
Ph: (310) 794-6685; Fax: (310) 267-4672

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