Skip to main content

Thank you for visiting nature.com. You are using a browser version with limited support for CSS. To obtain the best experience, we recommend you use a more up to date browser (or turn off compatibility mode in Internet Explorer). In the meantime, to ensure continued support, we are displaying the site without styles and JavaScript.

  • Protocol
  • Published:

Differential centrifugation–based biochemical fractionation of the Drosophila adult CNS

Nature Protocols volume 9, pages 2796–2808 (2014)Cite this article

Subjects

Abstract

Drosophila is widely used as a genetic model in questions of development, cellular function and disease. Genetic screens in flies have proven to be incredibly powerful in identifying crucial components for synapse formation and function, particularly in the case of the presynaptic release machinery. Although modern biochemical methods can identify individual proteins and lipids (and their binding partners), they have typically been excluded from use in Drosophila for technical reasons. To bridge this essential gap between genetics and biochemistry, we developed a fractionation method to isolate various parts of the synaptic machinery from Drosophila, thus allowing it to be studied in unprecedented biochemical detail. This is only possible because our protocol has unique advantages in terms of enriching and preserving endogenous protein complexes. The procedure involves decapitation of adult flies, homogenization and differential centrifugation of fly heads, which allow subsequent purification of presynaptic (and to a limited degree postsynaptic) components. It is designed to require only a rudimentary knowledge of biochemical fractionation, and it takes ∼3.5 h. The yield is typically 4 mg of synaptic membrane protein per gram of Drosophila heads.

This is a preview of subscription content, access via your institution

Access options

Buy this article

  • Purchase on Springer Link
  • Instant access to full article PDF
Buy now

Prices may be subject to local taxes which are calculated during checkout

Figure 1: Schematic workflow of the fly head fractionation with corresponding step numbers.
Figure 2: Representative illustrations from the fly head fractionation.
Figure 3: Analysis of subfractions taken during the procedure by SDS-PAGE and western blotting.

Similar content being viewed by others

References

  1. Lüscher, C. & Isaac, J.T. The synapse: center stage for many brain diseases. J. Physiol. 587, 727–729 (2009).

    Article  PubMed  PubMed Central  Google Scholar 

  2. Brenner, S. The genetics of Caenorhabditis elegans. Genetics 77, 71–94 (1974).

    CAS  PubMed  PubMed Central  Google Scholar 

  3. Suzuki, D.T., Grigliatti, T. & Williamson, R. Temperature-sensitive mutations in Drosophila melanogaster. VII. A mutation (parats) causing reversible adult paralysis. Proc. Natl. Acad. Sci. USA 68, 890–893 (1971).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  4. Bellen, H.J., Tong, C. & Tsuda, H. 100 years of Drosophila research and its impact on vertebrate neuroscience: a history lesson for the future. Nat. Rev. Neurosci. 11, 514–522 (2010).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  5. Richmond, J.E. & Broadie, K.S. The synaptic vesicle cycle: exocytosis and endocytosis in Drosophila and C. elegans. Curr. Opin. Neurobiol. 12, 499–507 (2002).

    Article  CAS  PubMed  Google Scholar 

  6. Chia, P.H., Li, P. & Shen, K. Cell biology in neuroscience: cellular and molecular mechanisms underlying presynapse formation. J. Cell Biol. 203, 11–22 (2013).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  7. Schoch, S. et al. Redundant functions of RIM1α and RIM2α in Ca2+-triggered neurotransmitter release. EMBO J. 25, 5852–5863 (2006).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  8. Südhof, T.C. The presynaptic active zone. Neuron 75, 11–25 (2012).

    Article  PubMed  PubMed Central  Google Scholar 

  9. Bronk, P. et al. Drosophila Hsc70-4 is critical for neurotransmitter exocytosis in vivo. Neuron 30, 475–488 (2001).

    Article  CAS  PubMed  Google Scholar 

  10. Hiesinger, P.R. et al. The v-ATPase V0 subunit a1 is required for a late step in synaptic vesicle exocytosis in Drosophila. Cell 121, 607–620 (2005).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  11. Lu, H., Leung, H.T., Wang, N., Pak, W.L. & Shieh, B.H. Role of Ca2+/calmodulin-dependent protein kinase II in Drosophila photoreceptors. J. Biol. Chem. 284, 11100–11109 (2009).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  12. Bachmann, A. et al. A perisynaptic ménage à trois between Dlg, DLin-7, and Metro controls proper organization of Drosophila synaptic junctions. J. Neurosci. 30, 5811–5824 (2010).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  13. Owald, D. et al. A Syd-1 homologue regulates pre- and postsynaptic maturation in Drosophila. J. Cell Biol. 188, 565–579 (2010).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  14. Nuwal, T., Heo, S., Lubec, G. & Buchner, E. Mass spectrometric analysis of synapsins in Drosophila melanogaster and identification of novel phosphorylation sites. J. Proteome Res. 10, 541–550 (2011).

    Article  CAS  PubMed  Google Scholar 

  15. Popescu, D.C., Ham, A.J. & Shieh, B.H. Scaffolding protein INAD regulates deactivation of vision by promoting phosphorylation of transient receptor potential by eye protein kinase C in Drosophila. J. Neurosci. 26, 8570–8577 (2006).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  16. Wagh, D.A. et al. Bruchpilot, a protein with homology to ELKS/CAST, is required for structural integrity and function of synaptic active zones in Drosophila. Neuron 49, 833–844 (2006).

    Article  CAS  PubMed  Google Scholar 

  17. Kittel, R.J. et al. Bruchpilot promotes active zone assembly, Ca2+ channel clustering, and vesicle release. Science 312, 1051–1054 (2006).

    Article  CAS  PubMed  Google Scholar 

  18. Fouquet, W. et al. Maturation of active zone assembly by Drosophila Bruchpilot. J. Cell Biol. 186, 129–145 (2009).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  19. Bodzon-Kulakowska, A. et al. Methods for samples preparation in proteomic research. J. Chromatogr. B Analyt. Technol. Biomed. Life Sci. 849, 1–31 (2007).

    Article  CAS  PubMed  Google Scholar 

  20. Huttner, W.B., Schiebler, W., Greengard, P. & De Camilli, P. Synapsin I (protein I), a nerve terminal-specific phosphoprotein. III. Its association with synaptic vesicles studied in a highly purified synaptic vesicle preparation. J. Cell Biol. 96, 1374–1388 (1983).

    Article  CAS  PubMed  Google Scholar 

  21. Ahmed, S., Holt, M., Riedel, D. & Jahn, R. Small-scale isolation of synaptic vesicles from mammalian brain. Nat. Protoc. 8, 998–1009 (2013).

    Article  PubMed  Google Scholar 

  22. Takamori, S. et al. Molecular anatomy of a trafficking organelle. Cell 127, 831–846 (2006).

    Article  CAS  PubMed  Google Scholar 

  23. Nagy, A., Baker, R.R., Morris, S.J. & Whittaker, V.P. The preparation and characterization of synaptic vesicles of high purity. Brain Res. 109, 285–309 (1976).

    Article  CAS  PubMed  Google Scholar 

  24. van de Goor, J., Ramaswami, M. & Kelly, R. Redistribution of synaptic vesicles and their proteins in temperature-sensitive shibirets1 mutant Drosophila. Proc. Natl. Acad. Sci. USA 92, 5739–5743 (1995).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  25. Kelly, L.E. The regulation of phosphorylation of a specific protein in synaptosomal fractions from Drosophila heads: the effects of light and two visual mutants. Cell Mol. Neurobiol. 3, 127–141 (1983).

    Article  CAS  PubMed  Google Scholar 

  26. Higashi-Kovtun, M.E., Mosca, T.J., Dickman, D.K., Meinertzhagen, I.A. & Schwarz, T.L. Importin-β11 regulates synaptic phosphorylated mothers against decapentaplegic, and thereby influences synaptic development and function at the Drosophila neuromuscular junction. J. Neurosci. 30, 5253–5268 (2010).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  27. Owald, D. et al. Cooperation of Syd-1 with Neurexin synchronizes pre- with postsynaptic assembly. Nat. Neurosci. 15, 1219–1226 (2012).

    Article  CAS  PubMed  Google Scholar 

  28. Whittaker, V.P., Michaelson, I.A. & Kirkland, R.J. The separation of synaptic vesicles from nerve-ending particles (′synaptosomes′). Biochem. J. 90, 293–303 (1964).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  29. Berninghausen, O. et al. Neurexin Iβ and neuroligin are localized on opposite membranes in mature central synapses. J. Neurochem. 103, 1855–1863 (2007).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  30. Gama Sosa, M.A., De Gasperi, R. & Elder, G.A. Modeling human neurodegenerative diseases in transgenic systems. Hum. Genet. 131, 535–563 (2012).

    Article  PubMed  Google Scholar 

  31. Okray, Z. & Hassan, B.A. Genetic approaches in Drosophila for the study neurodevelopmental disorders. Neuropharmacology 68, 150–156 (2013).

    Article  CAS  PubMed  Google Scholar 

  32. Karpinar, D.P. et al. Pre-fibrillar α-synuclein variants with impaired β-structure increase neurotoxicity in Parkinson′s disease models. EMBO J. 28, 3256–3268 (2009).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  33. Burré, J., Sharma, M. & Südhof, T.C. Systematic mutagenesis of α-synuclein reveals distinct sequence requirements for physiological and pathological activities. J. Neurosci. 32, 15227–15242 (2012).

    Article  PubMed  PubMed Central  Google Scholar 

  34. Neumüller, R.A. et al. Stringent analysis of gene function and protein-protein interactions using fluorescently tagged genes. Genetics 190, 931–940 (2012).

    Article  PubMed  PubMed Central  Google Scholar 

  35. Nadal, M.S. et al. The CD26-related dipeptidyl aminopeptidase-like protein DPPX is a critical component of neuronal A-type K+ channels. Neuron 37, 449–461 (2003).

    Article  CAS  PubMed  Google Scholar 

  36. Jerng, H.H., Kunjilwar, K. & Pfaffinger, P.J. Multiprotein assembly of Kv4.2, KChIP3 and DPP10 produces ternary channel complexes with ISA-like properties. J. Physiol. 568, 767–788 (2005).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  37. Boyken, J. et al. Molecular profiling of synaptic vesicle docking sites reveals novel proteins but few differences between glutamatergic and GABAergic synapses. Neuron 78, 285–297 (2013).

    Article  CAS  PubMed  Google Scholar 

  38. Dietzl, G. et al. A genome-wide transgenic RNAi library for conditional gene inactivation in Drosophila. Nature 448, 151–156 (2007).

    Article  CAS  PubMed  Google Scholar 

  39. Ni, J.Q. et al. A Drosophila resource of transgenic RNAi lines for neurogenetics. Genetics 182, 1089–1100 (2009).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  40. Bellen, H.J. et al. The BDGP gene disruption project: single transposon insertions associated with 40% of Drosophila genes. Genetics 167, 761–781 (2004).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  41. Rong, Y.S. et al. Targeted mutagenesis by homologous recombination in D. melanogaster. Genes Dev. 16, 1568–1581 (2002).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  42. Venken, K.J. & Bellen, H.J. Emerging technologies for gene manipulation in Drosophila melanogaster. Nat. Rev. Genet. 6, 167–178 (2005).

    Article  CAS  PubMed  Google Scholar 

  43. Beumer, K., Bhattacharyya, G., Bibikova, M., Trautman, J.K. & Carroll, D. Efficient gene targeting in Drosophila with zinc-finger nucleases. Genetics 172, 2391–2403 (2006).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  44. Carroll, D. Genome engineering with zinc-finger nucleases. Genetics 188, 773–782 (2011).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  45. Liu, J., Chen, Y. & Jiao, R. TALEN-mediated Drosophila genome editing: protocols and applications. Methods 69, 22–31 (2014).

    Article  CAS  PubMed  Google Scholar 

  46. Li, T. et al. Modularly assembled designer TAL effector nucleases for targeted gene knockout and gene replacement in eukaryotes. Nucleic Acids Res. 39, 6315–6325 (2011).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  47. Bassett, A.R. & Liu, J.L. CRISPR/Cas9 and genome editing in Drosophila. J. Genet. Genomics 41, 7–19 (2014).

    Article  CAS  PubMed  Google Scholar 

  48. Brand, A.H. & Perrimon, N. Targeted gene expression as a means of altering cell fates and generating dominant phenotypes. Development 118, 401–415 (1993).

    CAS  PubMed  Google Scholar 

  49. Frank, C.A. et al. New approaches for studying synaptic development, function, and plasticity using Drosophila as a model system. J. Neurosci. 33, 17560–17568 (2013).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  50. Dunkley, P.R., Jarvie, P.E. & Robinson, P.J. A rapid Percoll gradient procedure for preparation of synaptosomes. Nat. Protoc. 3, 1718–1728 (2008).

    Article  CAS  PubMed  Google Scholar 

  51. Peterson, G.L. A simplification of the protein assay method of Lowry et al. which is more generally applicable. Anal. Biochem. 83, 346–356 (1977).

    Article  CAS  PubMed  Google Scholar 

  52. Folin, O. & Ciocalteu, V. On tyrosine and tryptophane determinations in proteins. J. Biol. Chem. 73, 627–650 (1927).

    CAS  Google Scholar 

  53. Lowry, O.H., Rosebrough, N.J., Farr, A.L. & Randall, R.J. Protein measurement with the Folin phenol reagent. J. Biol. Chem. 193, 265–275 (1951).

    CAS  PubMed  Google Scholar 

  54. Nichols, R.A., Wu, W.C., Haycock, J.W. & Greengard, P. Introduction of impermeant molecules into synaptosomes using freeze/thaw permeabilization. J. Neurochem. 52, 521–529 (1989).

    Article  CAS  PubMed  Google Scholar 

  55. Begley, J.G. et al. Cryopreservation of rat cortical synaptosomes and analysis of glucose and glutamate transporter activities, and mitochondrial function. Brain Res. Brain Res. Protoc. 3, 76–82 (1998).

    Article  CAS  PubMed  Google Scholar 

  56. Daniel, J.A., Malladi, C.S., Kettle, E., McCluskey, A. & Robinson, P.J. Analysis of synaptic vesicle endocytosis in synaptosomes by high-content screening. Nat. Protoc. 7, 1439–1455 (2012).

    Article  CAS  PubMed  Google Scholar 

  57. VanBogelen, R.A. & Olson, E.R. Application of two-dimensional protein gels in biotechnology. Biotechnol. Annu Rev. 1, 69–103 (1995).

    Article  CAS  PubMed  Google Scholar 

  58. Matkovic, T. et al. The Bruchpilot cytomatrix determines the size of the readily releasable pool of synaptic vesicles. J. Cell Biol. 202, 667–683 (2013).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  59. Kaufmann, N., DeProto, J., Ranjan, R., Wan, H. & Van Vactor, D. Drosophila Liprin-α and the receptor phosphatase Dlar control synapse morphogenesis. Neuron 34, 27–38 (2002).

    Article  CAS  PubMed  Google Scholar 

  60. Liu, K.S. et al. RIM-binding protein, a central part of the active zone, is essential for neurotransmitter release. Science 334, 1565–1569 (2011).

    Article  CAS  PubMed  Google Scholar 

  61. Littleton, J.T., Bellen, H.J. & Perin, M.S. Expression of synaptotagmin in Drosophila reveals transport and localization of synaptic vesicles to the synapse. Development 118, 1077–1088 (1993).

    CAS  PubMed  Google Scholar 

  62. Mackler, J.M., Drummond, J.A., Loewen, C.A., Robinson, I.M. & Reist, N.E. The C2B Ca2+-binding motif of synaptotagmin is required for synaptic transmission in vivo. Nature 418, 340–344 (2002).

    Article  CAS  PubMed  Google Scholar 

  63. Zinsmaier, K.E., Eberle, K.K., Buchner, E., Walter, N. & Benzer, S. Paralysis and early death in cysteine string protein mutants of Drosophila. Science 263, 977–980 (1994).

    Article  CAS  PubMed  Google Scholar 

  64. Rubin, D.M. et al. Genomic structure and sequence analysis of Drosophila melanogaster HSC70 genes. Gene 128, 155–163 (1993).

    Article  CAS  PubMed  Google Scholar 

  65. Kirkpatrick, R.B. et al. Heavy chain dimers as well as complete antibodies are efficiently formed and secreted from Drosophila via a BiP-mediated pathway. J. Biol. Chem. 270, 19800–19805 (1995).

    Article  CAS  PubMed  Google Scholar 

  66. Khanna, M.R., Stanley, B.A. & Thomas, G.H. Towards a membrane proteome in Drosophila: a method for the isolation of plasma membrane. BMC Genomics 11, 302 (2010).

    Article  PubMed  PubMed Central  Google Scholar 

  67. Sun, B. & Salvaterra, P.M. Characterization of Nervana, a Drosophila melanogaster neuron-specific glycoprotein antigen recognized by anti–horseradish peroxidase antibodies. J. Neurochem. 65, 434–443 (1995).

    Article  CAS  PubMed  Google Scholar 

  68. Jacobs, H., Stratmann, R. & Lehner, C. A screen for lethal mutations in the chromosomal region 59AB suggests that bellwether encodes the α subunit of the mitochondrial ATP synthase in Drosophila melanogaster. Mol. Gen. Genet. 259, 383–387 (1998).

    Article  CAS  PubMed  Google Scholar 

  69. Ziviani, E., Tao, R.N. & Whitworth, A.J. Drosophila Parkin requires PINK1 for mitochondrial translocation and ubiquitinates Mitofusin. Proc. Natl. Acad. Sci. USA 107, 5018–5023 (2010).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  70. Celotto, A.M., Chiu, W.K., Van Voorhies, W. & Palladino, M.J. Modes of metabolic compensation during mitochondrial disease using the Drosophila model of ATP6 dysfunction. PLoS ONE 6, e25823 (2011).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  71. Kabani, M. & Martineau, C.N. Multiple Hsp70 isoforms in the eukaryotic cytosol: mere redundancy or functional specificity? Curr. Genomics 9, 338–348 (2008).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  72. Marchler, G. & Wu, C. Modulation of Drosophila heat shock transcription factor activity by the molecular chaperone DROJ1. EMBO J. 20, 499–509 (2001).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  73. Georgatos, S.D., Meier, J. & Simos, G. Lamins and lamin-associated proteins. Curr. Opin. Cell Biol. 6, 347–353 (1994).

    Article  CAS  PubMed  Google Scholar 

  74. Riemer, D. et al. Expression of Drosophila lamin C is developmentally regulated: analogies with vertebrate A-type lamins. J. Cell Sci. 108 (Part 10): 3189–3198 (1995).

    CAS  PubMed  Google Scholar 

  75. Sisson, J.C., Field, C., Ventura, R., Royou, A. & Sullivan, W. Lava lamp, a novel peripheral Golgi protein, is required for Drosophila melanogaster cellularization. J. Cell Biol. 151, 905–918 (2000).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

Download references

Acknowledgements

We thank R. Jahn for the opportunity to perform experiments in his laboratory at the Max Planck Institute for Biophysical Chemistry (MPIbpc) in Göttingen, Germany. Furthermore, we thank G. Mieskes for his technical advice regarding performance and documentation of experiments. We are grateful to H. Jäckle and members of his laboratory at the MPIbpc for use of their fly facilities. In particular, we thank R. Pflanz for his help in maintaining and expanding fly stocks. We thank M. Brünner (Freie Universität Berlin) for excellent technical assistance in purifying the custom polyclonal antibodies raised against Drosophila Liprin-α and RBP, as well as in performing the respective western blots. The monoclonal antibody ab49 (against DCSP-1) was developed by E. Buchner and A. Hofbauer and was obtained from the Developmental Studies Hybridoma Bank, created by the Eunice Kennedy Shriver National Institute of Child Health and Human Development (NICHD) of the US National Institutes of Health and maintained at the Department of Biology, The University of Iowa. This work was funded by grants from the Deutsche Forschungsgemeinschaft to S.J.S. (SFB 958 TP A6) and by a European Research Council Starting Grant (AstroFunc: 281961 to M.G.H.).

Author information

Authors and Affiliations

  1. Institute for Biology – Genetics, Freie Universität Berlin, Berlin, Germany

    Harald Depner, Janine Lützkendorf, Husam A Babkir & Stephan J Sigrist

  2. NeuroCure Cluster of Excellence, Charité, Berlin, Germany

    Stephan J Sigrist

  3. Laboratory of Glia Biology, Vlaams Instituut voor Biotechnologie (VIB) Center for the Biology of Disease, Katholieke Universiteit (KU) Leuven, Leuven, Belgium

    Matthew G Holt

Authors
  1. Harald Depner
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Janine Lützkendorf
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Husam A Babkir
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Stephan J Sigrist
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Matthew G Holt
    View author publications

    You can also search for this author in PubMed Google Scholar

Contributions

The experimental work was performed by H.D. and J. L., and analyzed by H.D., S.J.S. and M.G.H. H.A.B designed, cloned and purified the fusion protein construct used to generate the custom antibody against RBP that was used in this study. The project was conceived and supervised by M.G.H. and S.J.S. The manuscript was written by H.D., S.J.S and M.G.H., and commented on by all authors.

Corresponding author

Correspondence to Matthew G Holt.

Ethics declarations

Competing interests

The authors declare no competing financial interests.

Integrated supplementary information

Supplementary Figure 1 Specificity test of custom polyclonal antibodies used in this study.

Western blot analysis of adult fly head extracts probed with affinity-purified (a) anti-DRBPSH3 II+III and (b) anti-DLiprin-αC-term antibodies. (a) RBP expression analysis in control (rbp heterozygous animals: S201/+) and rbp mutant lines (rbpSTOP3/S201 and rbpMinos/S201). Two exposures of the same blot to light-sensitive autoradiography films are presented: 10 seconds (upper panel) and 30 seconds (middle panel). Bands of approximately 160 kDa and 190 kDa are detected (corresponding to the predicted sizes of RBP isoforms) in control animals only. An unspecific signal at around 130 kDa is also observed (most likely due to cross-reactivity of the anti-DRBPSH3 II+III antibody with other proteins in the fly head extract). (b) Liprin-α expression analysis in the indicated wild-type control (+/+ = w1118), heterozygous liprin-α animals (lipF3ex15/+) and liprin-α mutants (lipF3ex15/lipEPexR60). A single band around 140 kDa, corresponding to the predicted size of Liprin-α, is observed in control animals and is reduced in liprin-α heterozygous animals, with only faint signal in the hypomorphic liprin-α mutant lipF3ex15/lipEPexR60. α-Tubulin is displayed as a loading control for each western blot.

Supplementary information

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Depner, H., Lützkendorf, J., Babkir, H. et al. Differential centrifugation–based biochemical fractionation of the Drosophila adult CNS. Nat Protoc 9, 2796–2808 (2014). https://doi.org/10.1038/nprot.2014.192

Download citation

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1038/nprot.2014.192

This article is cited by

Comments

By submitting a comment you agree to abide by our Terms and Community Guidelines. If you find something abusive or that does not comply with our terms or guidelines please flag it as inappropriate.

Search

Advanced search

Quick links

Nature Briefing

Sign up for the Nature Briefing newsletter — what matters in science, free to your inbox daily.

Get the most important science stories of the day, free in your inbox. Sign up for Nature Briefing