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woensdag 8 augustus 2018

Nature Protocols Contents: Volume 13 Number 8

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Nature Protocols

August 2018 Volume 13, Issue 8

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A general, electrocatalytic approach to the synthesis of vicinal diamines    pp1725 - 1743
Niankai Fu, Gregory S. Sauer & Song Lin

1,2-diamines are commonly found in pharmaceutically relevant compounds. They can be prepared from diazides. This protocol describes an electrocatalytic method for preparing a wide range of diazides from alkenes, using a manganese catalyst.


Quantitative analysis of newly synthesized proteins    pp1744 - 1762
Yuanhui Ma, Daniel B. McClatchy, Salim Barkallah, William W. Wood & John R. Yates III

This protocol describes a proteomics approach for quantifying newly synthesized proteins (NSPs). NSPs are pulse-labeled with the methionine analog AHA or a heavy-isotope version of it, followed by AHA-peptide enrichment and LC-MS/MS analysis.


Assessing spatial pattern separation in rodents using the object pattern separation task    pp1763 - 1792
Nick P. van Goethem, Britt T. J. van Hagen & Jos Prickaerts

In this behavioral task, two objects are placed at specific locations at different times, and the response of mice and rats to these positions is assessed. The results indicate the rodents’ ability to recognize changes in spatial pattern separation.


Microfabrication of AngioChip, a biodegradable polymer scaffold with microfluidic vasculature    pp1793 - 1813
Boyang Zhang, Benjamin Fook Lun Lai, Ruoxiao Xie, Locke Davenport Huyer, Miles Montgomery et al.

This protocol describes the fabrication of AngioChip, a microfabricated scaffold for engineering vascularized tissues. In addition, the protocol contains procedures for immunostaining, drug screening and surgical anastomosis of the AngioChip.


Pig lung transplant survival model    pp1814 - 1828
Andrea Mariscal, Lindsay Caldarone, Jussi Tikkanen, Daisuke Nakajima, Manyin Chen et al.

This protocol describes a pig model in which lungs are transplanted into recipients who are subsequently monitored for 3 d after surgery to study ischemia reperfusion injury and early graft function.


Identification of RNA-binding protein targets with HyperTRIBE    pp1829 - 1849
Reazur Rahman, Weijin Xu, Hua Jin & Michael Rosbash

HyperTRIBE uses a hyperactive RNA-editing enzyme fused to an RNA-binding protein (RBP) to mark the target RNA transcripts of the RBP by converting adenosine to inosine near the binding sites with increased efficiency and reduced sequence bias.


Noninvasive 40-Hz light flicker to recruit microglia and reduce amyloid beta load    pp1850 - 1868
Annabelle C. Singer, Anthony J. Martorell, J. Miller Douglas, Fatema Abdurrob, Matthew K. Attokaren et al.

This protocol describes a noninvasive approach to evoke microglial engulfment and reduce amyloid levels in mouse brain. The authors describe assembly and operation of a light-flicker system, as well as assessment of the molecular and cellular effects.


Combined expansion microscopy with structured illumination microscopy for analyzing protein complexes    pp1869 - 1895
Yongfu Wang, Zulin Yu, Cori K. Cahoon, Tari Parmely, Nancy Thomas et al.

This protocol describes how to combine expansion microscopy (ExM) with structured illumination microscopy (SIM). ExM–SIM is exemplified by super-resolution analysis of the synaptonemal complex (SC) and single-particle averaging of SC proteins.


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