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#1
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| * HI! Avidin-Biotin interation is strongest known interaction till date. It has affinity constant of femtomolar range, followed by antigen antibody interaction which is of picomolar range on an average. You can try tagging protein A with Biotin and B with Avidin.It wont interfere with protein function if yours are plant proteins. Biotin is Vit.B2. hope it helps. Regards Shifali On Sat, 15 Nov 2008 Zhong Silin wrote : Shifali Chatrath Graduate Student Protein science Lab Dept. of Biological sciences National University of Singapore Singapore +65-65161210 +65-96393449 |
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#2
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| On Nov 16, 10:48*pm, "shifali chatrath" <[Only registered users see links. ]> wrote: Biotin is not a protein, and the OP was looking for an epitope tag to put on A. I don't know of an easy way to biotinylate one specific protein during biosynthesis. Also, vitamin B2 is riboflavin, not biotin. Nick -- Nick Theodorakis [Only registered users see links. ] contact form: [Only registered users see links. ] |
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#3
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| In article <[Only registered users see links. ]>, Nick Theodorakis <[Only registered users see links. ]> wrote: Biotinylation tag (a substrate peptide for E.coli biotin ligase) works quite well in E.coli (over 50% well-expressed protein modified) but does not work in eukariots. There is always an option to biotinylate purified protein in vitro using either purchased enzyme or expressing it yourself (which is easy because it's very soluble). When done right, in vitro biotynylated of the tagged protein is nearly 100% efficient. To the OP, I would suggest artificial coiled coils. Basically, a pair of 21 aa peptides used as interacting tags. One is very, very basic, another is very, very acidic - both conforming to the canonical coiled coil heptade repeat, so the only coiled coil formed is a heterodimer. I don't have a ref. but have a xerox somewhere at work. The affinity of such pair is subnanomolar. DK |
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#4
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| In <3kIUk.8581$[Only registered users see links. ]>, DK <[Only registered users see links. ]> wrote: I've used a modified version of the GCN4 leucine zipper for this purpose. Sadly, I don't have the reference handy - I am at home, but this worked very well to induce dimerization. If it helps any, the paper that I got the sequence from was from Peter Kim's lab, and published in Biochemistry. AC -- Email: echo 36434455860060025978157675027927670979097959886449 930P | dc |
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| interaction , proteinprotein , strongest |
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