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  2. → Viewing Profile: Posts: vh04x

vh04x

Member Since 26 Aug 2012
Offline Last Active Aug 31 2012 02:24 AM
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Posts I've Made

In Topic: Co-IP: Prey Protein Present in Neg and Pos Eluates

29 August 2012 - 07:40 AM

Thanks :)

The beads have aldehyde groups on them that react with primary amines to form Schiff bases. NaCNBH3 is then added to reduce the bond to a secondary amine. Any spare aldehyde groups are quenched with reacting with Tris.

It's all part of the Pierce Co-IP (Direct) Kit

In Topic: Co-IP: Prey Protein Present in Neg and Pos Eluates

28 August 2012 - 06:42 PM

Thanks for getting back to me.

No, none of ProA, B, C, or D have ZZ domains.

The Co-IP is performed from the lysates of platelets extracted from healthy donor blood. It's as physiological as it gets, I think.
Unfortunately, no one has managed to successfully make recombinant ProA, as it just falls apart when its synthesized out of the body.

ProA also has a nasty habit of binding to Protein A and Protein G, so this is Co-IP is performed with directly immobilized antibody (if that helps at all)

In Topic: PVDF membrane silver staining protocol

28 August 2012 - 06:35 PM

I'm confused. Why not just silver stain the gel before transfering to PVDF?

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