Welcome to Nature Protocols
Nature Protocols is an online resource for protocols, including authoritative, peer-reviewed 'Nature Protocols' and an interactive 'Protocols Network'. The two create a dynamic forum for scientists to upload and comment on protocols.
Featured Protocols
Genetic analysis
Positional cloning by fast-track SNP-mapping in Drosophila melanogasterPositional cloning of chemically-induced mutations in Drosophila can take many years using traditional methods. Here Schnorrer et al. describe an SNP-based approach that can determine the chromosomal location of mutations with a resolution of 50kb -- all in a matter of months.
Genomics/proteomics
The SCX/IMAC enrichment approach for global phosphorylation analysis by mass spectrometryMass spectrometry(MS)-based proteomics methods can struggle to detect phosphoproteins because they tend to be expressed and phosphorylated at low levels. This protocol describes how to enrich the proteome for phosphoproteins based on their charge (using strong cation exchange, SCX) and on their affinity to certain metal ions (using immobilized metal affinity chromatography, IMAC). Individual phosphoproteins within the enriched samples can be subsequently identified by LC-MS/MS
IMAGING
Labeling and visualization of blood vessels with DiI dyeThis protocol provides a method for labeling blood vessels using the lipophilic carbocyanine dye DiI, which enables visualization of vasculature in experimental animals by conventional and confocal fluorescence microscopy.
Genetic Analysis
Blocking miRNA function with antisense oligonucleotides (ASOs)MicroRNAs (miRNAs) are involved in gene regulation, but assigning specific functions to individual miRNAs has been problematic. This protocol from the Zamore lab describes how to design ASOs to block the function of specific miRNAs and how to deliver these ASOs efficiently to cultured Drosophila or mammalian cells.
Biochemistry and Protein Analysis
Absolute quantitation of intracellular metabolite concentrations Quantitation of cellular metabolic fluxesThese are two protocols for quantifying metabolite concentrations in cultured cells (mammalian or E. coli) using 13C-labeled carbon sources followed by analysis by LC-MS.
Protocols Network
There are various ways you can contribute:
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Highlights from the network
- Identification and analysis of mutated ALK in Neuroblastoma
- Electrophysiology Measurement of Long-term Plasticity of Developing GABAergic Synapses in Acute Rat Hippocampal Slices.
- Subunit stoichiometry determination by chemical cross-linking
- Subunit stoichiometry determination by perfluorooctanoic acid polyacrylamide gel electrophoresis (PFO-PAGE)
- Production of neuron-preferential lentiviral vectors
- Isolation of murine hepatocytes to measure protein synthesis, ex vivo, upon stimulation
- AlignDB: A computational protocol for analyzing the relationship between indels and SNPs in genome sequences
- Novel mouse glioma model: Cell-type and region specific activation of oncogenes using lentiviral vectors
- Cellular transformation and leukemia development by activated tyrosine kinases
- In situ Protocol for embryos and juveniles of Convolutriloba longifissura
- Quantile Regression for Trend Analysis
- Isolation of murine lamina propria CD11c+ cells
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