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Method Article
Thomas Gilmore
Gilmore lab, Boston University
Corresponding Author
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Among basal animals, the starlet sea anemone Nematostella vectensis (phylum Cnidaria) has emerged as a leading laboratory model organism. Herein, we describe techniques to prepare antiserum against Nematostella proteins and to detect Nematostella proteins by Western blotting of extracts from animals or tissue culture cells overexpressing Nematostella cDNAs. These protocols may also be used to complement a variety of cellular and molecular techniques being used by the increasing number of investigators who are focusing on gene regulatory systems of basal animals, such as sponges, corals, and placozoans.
Abbreviations: aa, amino acid(s); ASW, artificial sea water; BSA, bovine serum albumin; EST, expressed sequence tag; HRP, horseradish peroxidase; KLH, Keyhole Limpet hemocyanin; NGS, normal goat serum; Nv, Nematostella vectensis; PBS, phosphate-buffered saline; SDS, Sodium dodecyl sulfate; TBS, Tris-buffered saline; TBST, Tris-buffered saline-Tween-20.
Keywords
Nematostella; antiserum; Western blotting
Figure 1
The authors declare no competing financial interests.
This is a list of supplementary files associated with this preprint. Click to download.
- supplement0.doc
Table 1 Antisera used for Western blotting.
- supplement0.doc
Table 2 Troubleshooting table.
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Subject Areas
Cell biology
Developmental biology
Computational biology and bioinformatics
Biological techniques
Associated Publications
Characterization of the Core Elements of the NF- kappaB Signaling Pathway of the Sea Anemone Nematostella vectensis
by F. S. Wolenski, M. R. Garbati, T. J. Lubinski, +6
Molecular And Cellular Biology (25 November, 2012)
Characterizing the spatiotemporal expression of RNAs and proteins in the starlet sea anemone, Nematostella vectensis
by Francis S Wolenski, Michael J Layden, Mark Q Martindale, +2
Nature Protocols (18 April, 2013)
An open repository of community-contributed protocols sponsored by Nature Research.
Learn more about Protocol Exchange
Method Article
Thomas Gilmore
Gilmore lab, Boston University
Corresponding Author
Version History
CURRENT STATUS: Posted
Version 1
Posted 13 Dec, 2012
No community comments so far
Metrics
Comments: 0
HTML Views: ...
License:
This work is licensed under a CC BY-NC 3.0 License. Read Full License
Among basal animals, the starlet sea anemone Nematostella vectensis (phylum Cnidaria) has emerged as a leading laboratory model organism. Herein, we describe techniques to prepare antiserum against Nematostella proteins and to detect Nematostella proteins by Western blotting of extracts from animals or tissue culture cells overexpressing Nematostella cDNAs. These protocols may also be used to complement a variety of cellular and molecular techniques being used by the increasing number of investigators who are focusing on gene regulatory systems of basal animals, such as sponges, corals, and placozoans.
Abbreviations: aa, amino acid(s); ASW, artificial sea water; BSA, bovine serum albumin; EST, expressed sequence tag; HRP, horseradish peroxidase; KLH, Keyhole Limpet hemocyanin; NGS, normal goat serum; Nv, Nematostella vectensis; PBS, phosphate-buffered saline; SDS, Sodium dodecyl sulfate; TBS, Tris-buffered saline; TBST, Tris-buffered saline-Tween-20.
Figure 1
The authors declare no competing financial interests.
This is a list of supplementary files associated with this preprint. Click to download.
- supplement0.doc
Table 1 Antisera used for Western blotting.
- supplement0.doc
Table 2 Troubleshooting table.
Loading...
Associated Publications
Characterization of the Core Elements of the NF- kappaB Signaling Pathway of the Sea Anemone Nematostella vectensis
by F. S. Wolenski, M. R. Garbati, T. J. Lubinski, +6
Molecular And Cellular Biology (25 November, 2012)
Characterizing the spatiotemporal expression of RNAs and proteins in the starlet sea anemone, Nematostella vectensis
by Francis S Wolenski, Michael J Layden, Mark Q Martindale, +2
Nature Protocols (18 April, 2013)