Sybody Generation Toolbox
(Kit #
1000000160
)
Depositing Lab: Markus Seeger
The Seeger Lab has established a protocol to select synthetic nanobodies – called sybodies – against (membrane) protein targets. This collection contains all plasmids necessary for the sybody generation process.
This kit will be sent as bacterial glycerol stocks in 96-well plate format.
Original Publications
Generation of Synthetic Nanobodies Against Delicate Proteins. Zimmermann I, Egloff P, Hutter CAJ, Kuhn BT, Bräuer P, Newstead S, Dawson RJP, Geertsma ER, Seeger MA. Nature Protocols. 2020 Apr 8. doi: 10.1038/s41596-020-0304-x. PubMed PMID: 32269381.
Synthetic Single Domain Antibodies for the Conformational Trapping of Membrane Proteins. Zimmermann I, Egloff P, Hutter CA, Arnold FM, Stohler P, Bocquet N, Hug MN, Huber S, Siegrist M, Hetemann L, Gera J, Gmür S, Spies P, Gygax D, Geertsma ER, Dawson RJ, Seeger MA. Elife. 2018 May 24;7. pii: 34317. doi: 10.7554/eLife.34317. PubMed PMID: 29792401.
Description
The Seeger Lab has established a general protocol to select synthetic nanobodies – called sybodies – against soluble proteins, membrane proteins, and protein complexes. This collection contains all plasmids necessary for the entirely in vitro sybody generation process. A detailed protocol referencing all of the plasmids can be found in the Nature Protocols article.
- Schematic overview of the sybody platform. Sybodies against protein targets can be generated within three weeks.
Protocols
Generation of Synthetic Nanobodies Against Delicate Proteins. Zimmermann I, Egloff P, Hutter CAJ, Kuhn BT, Bräuer P, Newstead S, Dawson RJP, Geertsma ER, Seeger MA. Nature Protocols. 2020 Apr 8. doi: 10.1038/s41596-020-0304-x. PubMed PMID: 32269381.
How to Cite this Kit
These plasmids were created by your colleagues. Please acknowledge the Principal Investigator, cite the article in which they were created, and include Addgene in the Materials and Methods of your future publications.
For your Materials and Methods section:
“The Sybody Generation Toolbox was a gift from Markus Seeger (Addgene kit #1000000160).”
For your Reference section:
Generation of Synthetic Nanobodies Against Delicate Proteins. Zimmermann I, Egloff P, Hutter CAJ, Kuhn BT, Bräuer P, Newstead S, Dawson RJP, Geertsma ER, Seeger MA. Nature Protocols. 2020 Apr 8. doi: 10.1038/s41596-020-0304-x. PubMed PMID: 32269381.
Synthetic Single Domain Antibodies for the Conformational Trapping of Membrane Proteins. Zimmermann I, Egloff P, Hutter CA, Arnold FM, Stohler P, Bocquet N, Hug MN, Huber S, Siegrist M, Hetemann L, Gera J, Gmür S, Spies P, Gygax D, Geertsma ER, Dawson RJ, Seeger MA. Elife. 2018 May 24;7. pii: 34317. doi: 10.7554/eLife.34317. PubMed PMID: 29792401.
Sybody Generation Toolbox - #1000000160
- Resistance Color Key
Each circle corresponds to a specific antibiotic resistance in the kit plate map wells.
- Inventory
Searchable and sortable table of all plasmids in kit. The Well column lists the plasmid well location in its plate. The Plasmid column links to a plasmid's individual web page.
- Kit Plate Map
96-well plate map for plasmid layout. Hovering over a well reveals the plasmid name, while clicking on a well opens the plasmid page.
Resistance Color Key
| Chloramphenicol and Ampicillin | |
| Chloramphenicol | |
| Ampicillin |
Inventory
| Well | Plasmid | Resistance |
|---|---|---|
| A / 1 | pBXNPH3 | Chloramphenicol and Ampicillin |
| A / 2 | pBXNPHM3 | Chloramphenicol and Ampicillin |
| A / 3 | pSBinit | Chloramphenicol |
| A / 4 | pSb_init containing the TM287/288 nanobody Nb_TM#2 | Chloramphenicol |
| A / 5 | pSb_init containing the MBP sybody Sb_MBP#1 | Chloramphenicol |
| A / 6 | pDXinit | Ampicillin |
| A / 7 | SB_concave | Ampicillin |
| A / 8 | SB_loop | Ampicillin |
| A / 9 | SB_convex | Ampicillin |
| A / 10 | pRDV5 containing Sb-loop | Ampicillin |
| A / 11 | pRDV5 containing Sb-convex | Ampicillin |
| A / 12 | pDX_init containing Sb-loop | Ampicillin |
| B / 1 | pBXNH3CA_MBP | Ampicillin |
| B / 2 | pBXNH3LCA_TM287/288 | Ampicillin |