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Method Article
Preparation of antiserum and detection of proteins by Western blotting using the starlet sea anemone, Nematostella vectensis
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
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Table 1 Antisera used for Western blotting.
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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)
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This is a preprint. It has not completed peer review.
Method Article
Preparation of antiserum and detection of proteins by Western blotting using the starlet sea anemone, Nematostella vectensis
Thomas Gilmore
Gilmore lab, Boston University
Corresponding Author
Badges
Prescreen
This manuscript passed our Prescreen assessment
Complete author information
Appropriate declaration statements
Potential risks to human health
Prescreen
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
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)