fractions Protocols

Search results for: fractions

Protocols » Search Results

Search Results Protocol Links Sort by: Hits | Alphabetical

CHO Centrosome Prep Protocol - http://mitchison.med.harvard.edu/protocols/mt1.html

Category: Cell Organelle Protocols: Nucleus Protocols

The key step is the lysis which solubilizes centrosomes away from nuclei by very low ionic strength lysis after treatment of cells with nocodazole and cytochalasin B. The released centrosomes are then centrifuged onto a Ficoll cushion (to avoid pelleting) and the interface between the lysate and the Ficoll is collected and the centrosomes are concentrated on a sucrose gradient. Fractions are assayed by spindown and double IF with 5051 serum and anti-tubulin and the pooled fractions are frozen... - [Read CHO Centrosome Prep Protocol]

Isolation of Arabidopsis Nuclei and Measurement of Gene Transcription Rates Protocol - http://www.natureprotocols.com/2007/01/25/isolation_of_arabidopsis_nucle.php

Category: Plant Biology Protocols: Arabidopsis Genetics Protocols

Protocol for the isolation of Arabidopsis nuclei and measurement of gene transcription rates using nuclear run-on assays. Plant materials are ground in hexylene glycol-based buffers and highly enriched nuclear fractions are obtained using Percoll density gradients. Standard and small-scale protocols are presented, along with a tested method for nuclear run-on assays. The entire process may be completed within 3 days. - [Read Isolation of Arabidopsis Nuclei and Measurement of Gene Transcription Rates Protocol]

Lipid Extraction from Plasma Membrane Fractions Prepared from Yeast Cells Protocol - http://www.bio.com/protocolstools/protocol.jhtml?id=p419

Category: Lipid Protocols

Protocol for lipid extraction from plasma membrane fractions prepared from yeast cells. - [Read Lipid Extraction from Plasma Membrane Fractions Prepared from Yeast Cells Protocol]

Neuron Culture Solutions - http://www.ehlerslab.org/protocols.html

Category: Neuroscience: Neuron Cell Culture Protocols

Neuron Cell Culture Solutions, Neuron Culturing, Preparation of Brain Membrane Fractions, Biotinylation Assay of Receptor Endocytosis, PSD Prep. Ehlers lab Duke Univ. - [Read Neuron Culture Solutions]

Northern Blot Analysis of mRNA From Mammalian Polyribosomes Protocol - http://www.natureprotocols.com/2006/09/15/northern_blot_analysis_of_mrna.php

Category: Nucleic Acid Detection Protocols: Nucleic Acid Blotting Protocols

The protocol consists of a method for the generation of cytoplasmic extracts from mammalian cells (in this case, 293T cells) without the disruption of polyribosomes, the separation of ribosomal components and polyribosomes by sucrose gradient centrifugation, the isolation of mRNA from these fractions, and detection of mRNA by Northern blot analysis. - [Read Northern Blot Analysis of mRNA From Mammalian Polyribosomes Protocol]

Preparation of Arabidopsis Chloroplasts Protocol - http://www.cshprotocols.org/cgi/content/abstract/2007/1/pdb.prot4681

Category: Plant Biology Protocols: Arabidopsis Tissue Culture

It is desirable to prepare subcellular fractions, either to localize proteins or to improve the sensitivity of protein detection. This procedure describes the enrichment of chloroplasts from Arabidopsis. - [Read Preparation of Arabidopsis Chloroplasts Protocol]

Preparation of Arabidopsis Mitochondria Protocol - http://www.cshprotocols.org/cgi/content/abstract/2007/1/pdb.prot4682

Category: Plant Biology Protocols: Arabidopsis Tissue Culture

It is often desirable to prepare subcellular fractions, either to localize proteins or to improve the sensitivity of protein detection. This procedure describes the enrichment of mitochondria from Arabidopsis. - [Read Preparation of Arabidopsis Mitochondria Protocol]

Preparation of Brain Membrane Fractions for Western Blot Analysis - http://www.bdbiosciences.com/pharmingen/protocols/Brain_Membrane_Fractions.shtml

Category: Protein Protocols: Western Blot Protocol

Western blotting using enriched membrane preparation and postsynaptic densities are desired. (BD Biosciences) - [Read Preparation of Brain Membrane Fractions for Western Blot Analysis]

Preparation of Crude Subcellular Fractions by Differential Centrifugation Protocol - http://www.axis-shield.com/densityhome/optiprep/S07.pdf

Category: Cell Organelle Protocols: Subcellular Fractionation Protocols

The employment of differential centrifugation to prepare crude fractions of subcellular particles from homogenates is often a necessary first step to a subsequent purification of one or more particles on a density gradient. This protocol describes the use of differential centrifugation to fractionate a mammalian liver homogenate but similar methods should be applicable to all mammalian tissues and cultured cells. - [Read Preparation of Crude Subcellular Fractions by Differential Centrifugation Protocol]

Preparing Vacuolar Membranes And Other Membrane Fractions from Neurospora Crassa Protocol - http://www.fgsc.net/neurosporaprotocols/VM%20vacuole%20procedure.pdf

Category: Fungi Protocols: Neurospora Protocols

Protocol for Preparing Vacuolar Membranes And Other Membrane Fractions from Neurospora Crassa (4 liter prep, easily scaled up to 10, 12, 24, or 30 l). - [Read Preparing Vacuolar Membranes And Other Membrane Fractions from Neurospora Crassa Protocol]

Purification of leukocyte fractions from rat peritoneal exudates by density barrier sedimentation. - http://www.axis-shield.com/densityhome/optiprep/C19.pdf

Category: Cell Biology and Cell Culture

The separation depends on the use of a hyperosmotic density barrier to separate the two types of leukocyte. - [Read Purification of leukocyte fractions from rat peritoneal exudates by density barrier sedimentation.]

Recycling Tubulin Protocol - http://mitchison.med.harvard.edu/protocols/recycle.html

Category: Cytoskeleton Protocols: Microtubules Protocols

Recycle tubulin fractions stored at -80¡C after the PC column and store the recycled tubulin in small aliquots for day-to-day use. Generally store recycled tubulin in Injection Buffer (IB) without free GTP. This is done because depolymerization appears to be much better in IB, IB is ideal for microinjections/adding tubulin to extracts, and the absence of free GTP makes polymerization with GMPCPP, a very useful GTP analog that has ~5-10X lower affinity than GTP for tubulin. - [Read Recycling Tubulin Protocol]

Tissue subcellular fractionation and protein extraction for use in mass-spectrometry-based proteomic - http://www.nature.com/nprot/journal/v1/n4/abs/nprot.2006.273.html

Category: Protein Protocols: Protein Extraction From Tissue

Tissue subcellular fractionation and protein extraction for use in mass-spectrometry-based proteomics. Although many methods exist for fractionating proteins, the method described here can capture the majority of subcellular fractions simultaneously at reasonable purity. The scalability of this method makes it amenable to small samples, such as embryonic tissues, in addition to larger tissues. The protocol described is for the general fractionation and extraction of proteins from organs / tissue - [Read Tissue subcellular fractionation and protein extraction for use in mass-spectrometry-based proteomic]