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Linkage chemistries
- OldForum
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12 years 7 months ago #1
by OldForum
Linkage chemistries was created by OldForum
How is carboxymethyl dextran covalently coupled to the gold surface? What crosslinking reagent is used?
jason
jason
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- OldForum
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12 years 7 months ago - 9 years 9 months ago #2
by OldForum
Replied by OldForum on topic Linkage chemistries
Hi Jason,
The dextran is linked to the gold with a SAM (self assebled monolayer).
see: www.sprpages.nl/spr-overview/sensor-chips.html
The dextran is linked to the gold with a SAM (self assebled monolayer).
see: www.sprpages.nl/spr-overview/sensor-chips.html
Last edit: 9 years 9 months ago by .
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- OldForum
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12 years 7 months ago #3
by OldForum
Replied by OldForum on topic Linkage chemistries
Hi. Yes, thank you but I'm looking for the specific protocol. I can link a protein to gold with DSP. 11-mercaptododecanol is commonly used as a SAM. An epoxide can be used to link the SAM to the sugar but how do I make sure the dextran preferentially binds to the SAM and does not crosslink with itself?
Thanks
jason
Thanks
jason
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- OldForum
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12 years 7 months ago #4
by OldForum
Replied by OldForum on topic Linkage chemistries
Mmm, I don't have a specific protocol.
Dextran has free -COOH groups at the end of the chains. My hunch would be that the internal cross-linking between the dextran is not a real issue when you use chemistries targetting the -COOH groups.
see: en.wikipedia.org/wiki/Dextran
Arnoud
Dextran has free -COOH groups at the end of the chains. My hunch would be that the internal cross-linking between the dextran is not a real issue when you use chemistries targetting the -COOH groups.
see: en.wikipedia.org/wiki/Dextran
Arnoud
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- OldForum
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12 years 7 months ago #5
by OldForum
Replied by OldForum on topic Linkage chemistries
Dextran must be carboxylated with bromoacetic acid right? I've found this paper so far...http://www.sciencedirect....le/pii/S0956566302000842
Another option might be to create aldehydes on the dextran and link that to hydrazide.
jason
Another option might be to create aldehydes on the dextran and link that to hydrazide.
jason
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12 years 7 months ago #6
by OldForum
Replied by OldForum on topic Linkage chemistries
Hi Jason, yes thats right: I use 1 M in 2 N NaOH overnight in a humidity chamber.
Be careful because the 2 N NaOH will etch (in case of Biacore chips, do not let it touch the plastic and glue!
The ref I use is:
Chatelier, R. C., et al. "A general method to recondition and reuse BIAcore sensor chips fouled with covalently immobilized protein/peptide." Analytical Biochemistry 229.1 (1995): 112-18.
Arnoud
Be careful because the 2 N NaOH will etch (in case of Biacore chips, do not let it touch the plastic and glue!
The ref I use is:
Chatelier, R. C., et al. "A general method to recondition and reuse BIAcore sensor chips fouled with covalently immobilized protein/peptide." Analytical Biochemistry 229.1 (1995): 112-18.
Arnoud
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