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 help with my glutathione sepharose [View Printable]
spanishflower

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hello everybody
i am working on a GST taged protein and when i do the glutathione sepharose beads binding assay i found difficulty in the binding?
i dont know why?
i am using the Batch method and did add 10 mMDDT to the lysis buffer and increased the incubation time for binding to 2 hours in 4 degree.......
dont know whaat else to do
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 Posted Jun 23, 2006, 2:18 AM
spanishflower

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help anybody????
.........................
Posted Jun 28, 2006, 4:56 AM
gsovak

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Hi,
I never did it with GST and Gluthatione.
I did with HA tagged and anti HA ab.
I found this article maybe you can try their protocol.
Arndt V, Daniel C, Nastainczyk W, Alberti S, Hohfeld J. Related Articles, Links
BAG-2 acts as an inhibitor of the chaperone-associated ubiquitin ligase CHIP.
Mol Biol Cell. 2005 Dec;16(12):5891-900. Epub 2005 Oct 5.
PMID: 16207813 [PubMed - indexed for MEDLINE]
.........................
Posted Jun 28, 2006, 18:35 PM
spanishflower

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THANKXXXXXXX
.........................
Posted Jun 29, 2006, 2:08 AM
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