|Item||Catalog #||Description||Quantity||Price (USD)|
|Plasmid||36099||Standard format: Plasmid sent in bacteria as agar stab||1||$75|
This material is available to academics and nonprofits only.
- Backbone size (bp) 5450
Modifications to backboneBT-tag (biotinilation site from P.freudenreichia transcarboxilase, separated from MCS by TEV-site)
Vector typeMammalian Expression
- Promoter EF1a
/ Fusion Protein
- BT-tag (C terminal on insert)
Growth in Bacteria
Copy numberHigh Copy
- Cloning method Restriction Enzyme
- 5′ sequencing primer CTCAAGCCTCAGACAGTGG
- 3′ sequencing primer GAGGGGTGCAAGGACCCAG (Common Sequencing Primers)
After mammal transfection, supplement fresh media with 4 uM Biotin to "charge" tag by Biotin. Allow cells to grow for about 48 h before the purification steps.
These plasmids were created by your colleagues. Please acknowledge the Principal Investigator, cite the article in which the plasmids were described, and include Addgene in the Materials and Methods of your future publications.
For your Materials & Methods section:pEBB-TB was a gift from Ezra Burstein (Addgene plasmid # 36099 ; http://n2t.net/addgene:36099 ; RRID:Addgene_36099)
For your References section:A bimolecular affinity purification method under denaturing conditions for rapid isolation of a ubiquitinated protein for mass spectrometry analysis. Maine GN, Li H, Zaidi IW, Basrur V, Elenitoba-Johnson KS, Burstein E. Nat Protoc. 2010 Aug;5(8):1447-59. Epub 2010 Jul 22. 10.1038/nprot.2010.109 PubMed 20671728