Protocols

Below is a working list of NCDIR protocols and methodologies. Please contact us if you have questions or comments, or would like to submit a protocol.  

Disruption of Yeast or Bacteria Cells

Harvesting Cells and Making Noodles

Harvesting Cells and Making Noodles (video protocol)

Cryogenic Disruption of Yeast Cells PM100

Cryogenic Disruption Using PM100 Ball Mill (video protocol)

Microfluidizer

Zymolase Treatment  

Disruption of Mammalian Cells

Cryogenic Disruption of Mammalian Cells using Retsch PM 100

Protein Affinity Capture from Cryogenically Disrupted Human Cells  

Subcellular Fractionation

Isolation of Yeast Nuclear Envelopes and Nuclear Pore Complex 

Preparation of Yeast Nuclei & Spindle Pole Bodies  Carbonate

Extraction of Nuclear Envelopes Rapid, Small-Scale Subcellular

Fractionation for Nuclei and Nuclear Envelopes Heparin Extraction of Nuclear Envelopes  

Isolation of Protein Complexes

Multi-well Interactomic Screen Affinity proteomics to study endogenous protein complexes

Conjugation of Tosylactivated Dynabeads

Conjugation of Magnetic Beads

Pullouts with IgG Resin

Immunoaffinity purifications of protein complexes  

Mass Spectrometry

MALDI Sample Preparation_In-gel Digestion and Crystallization

MALDI Sample Preparation_The Ultra Thin Layer Method MALDI

Sample Preparation_Saturated 4-HCCA Solution

IMAC phosphopeptide enrichment Phosphopeptide Enrichment Using TiO2 GL Biosciences  

Genomic Analyses

mRNA Expression Microarrays Agilent Expression Microarrays ChIP (chromatin immunoprecipitation) for ChIP-CHIP analysis

Yeast Quantitative PCR Protocol  

Protein Biochemistry

Carbonate Silver Stain

Electropurification

Methanol Precipitation of Protein

Protease Mapping

Strip Blots

Whole Cell Lysis and TCA (Trichloroacetic acid) Precipitation