lower Protocols

Search results for: lower

Protocols » Search Results

Search Results Protocol Links Sort by: Hits | Alphabetical

Long-PCR Reagents and Guidelines - http://wheat.pw.usda.gov/~lazo/methods/lazo/longpcr.html

Category: PCR Protocols: Quantitative PCR Protocols

General guidelines for long-PCR conditions and enzyme mixtures. Efficient long-PCR results from the use of two polymerases: a non-proofreading polymerase is the main polymerase in the reaction, and a proofreading polymerase (3' to 5' exo) is present at a lower concentration. Includes: For PCR with low-complexity templates (e.g., plasmid and cosmid inserts); For PCR with moderate-complexity templates (e.g., bacterial genomic DNA); For PCR with high-complexity templates (e.g., human genomic DNA). - [Read Long-PCR Reagents and Guidelines]

Preparation of Electrolyte Gradient Gels Protocol - http://www.cshprotocols.org/cgi/content/extract/2006/2/pdb.prot3795

Category: Nucleic Acid Electrophoresis Protocols

Protocol for the preparation of electrolyte gradient gels. Electrolyte gradients are formed when buffers of different concentrations are used in the upper (low electrolyte concentration) and lower (high electrolyte concentration) chambers of the electrophoresis device. - [Read Preparation of Electrolyte Gradient Gels Protocol]

Protocol for L-selectin-PNAd Interactions under Flow Conditions. - http://www.cbrinstitute.org/labs/springer/protocols/azucena_flow_chamber.html

Category: Cell Biology and Cell Culture

This protocol focuses on the interactions between L-selectin expressed on neutrophils and PNAd coated onto the plastic surface. The main purpose of the flow chamber assay is to visualize and measure interactions between flowing cells expressing a given adhesion molecule on their surface, and their receptor, either directly coated on the flow chamber lower wall or expressed on a cell monolayer. - [Read Protocol for L-selectin-PNAd Interactions under Flow Conditions.]

Purification of Demethylated Sphingomyelin Protocol - http://www.duke.edu/web/ceramide/protocols/0032.html

Category: Lipid Protocols

Protocol for the purificiation of demethylated sphingomyelin. Includes: Lower, chloroform phase; Upper, aqueous phase and interphase. - [Read Purification of Demethylated Sphingomyelin Protocol]

Purification of Histidine-Tagged Proteins Using IMAC Without Parameter Optimization Protocol - http://www.cshprotocols.org/cgi/content/extract/2006/1/pdb.prot4217

Category: Chromatography Protocols: Immobilized Metal Ion Affinity Chromatography Protocols

Histidine-tagged proteins can be purified on prepacked 1-ml immobilized metal-ion affinity chromatography (IMAC) columns without optimization of the separation conditions. The method allows fast capture of the target protein, although with a lower purity than can be obtained under optimized conditions. - [Read Purification of Histidine-Tagged Proteins Using IMAC Without Parameter Optimization Protocol]

Purification of Peroxisomes using a Density Barrier in a Swinging-Bucket Rotor - http://www.axis-shield.com/densityhome/optiprep/S10.pdf

Category: Cell Organelle Protocols: Subcellular Fractionation Protocols: Peroxisome Isolation Protocols

Peroxisomes can be purified in iodixanol gradients in high yield (80-90%) with no detectable contamination from any other organelle. This is a property unique to iodixanol because the densities of other organelles, particularly that of mitochondria (approx ρ = 1.14 g/ml) and endoplasmic reticulum (approx ρ = 1.13 g/ml) are much lower than that of peroxisomes (approx ρ = 1.18 g/ml). - [Read Purification of Peroxisomes using a Density Barrier in a Swinging-Bucket Rotor]

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]

Transformation of Agrobacterium Using Electroporation Protocol - http://www.cshprotocols.org/cgi/content/abstract/2006/30/pdb.prot4665

Category: Bacteria Transformation Protocols

Protocol describes a method for transforming Agrobacterium with plasmid DNA using electroporation in a manner similar to that commonly used for Escherichia coli. Although the transformation efficiency for Agrobacterium is lower than that for E. coli, it is possible to obtain adequate numbers of Agrobacterium transformants with this technique. - [Read Transformation of Agrobacterium Using Electroporation Protocol]

Transformation of Agrobacterium Using the Freeze-Thaw Method Protocol - http://www.cshprotocols.org/cgi/content/abstract/2006/30/pdb.prot4666

Category: Bacteria Transformation Protocols

Protocol describes a method for transforming Agrobacterium with plasmid DNA using a freeze-thaw technique. Although the transformation efficiency for Agrobacterium is lower than that for Escherichia coli, it is possible to obtain adequate numbers of transformants with this technique. - [Read Transformation of Agrobacterium Using the Freeze-Thaw Method Protocol]

TransWell Chemotaxis Protocol. - http://bioprotocol.bio.com/protocolstools/protocol.jhtml?id=p2156

Category: Immunology: Chemotaxis

TransWell Chemotaxis protocol. trans-well chemotaxis method from bioprotocol. The protocol is a method for studying migration towards a concentration gradient of chemoattractant of leukocytes (neutrophils, monocytes and lymphocytes) or other migratory cells. An upper chamber containing a suspension of cells is separated by a membrane from a lower chamber containing medium with chemoattractant. Chemotaxis of the cells from the upper chamber into the lower chamber can be quantified. - [Read TransWell Chemotaxis Protocol.]

Troubleshoot Immunoprecipitation - http://www.chemicon.com/resource/ANT101/a2.asp#TroubleShooting

Category: Protein Protocols: Immunoprecipitation Protocols: Immunoprecipitation Troubleshooting

Troubleshoot Immunoprecipitation. Chemicon. The most common challenge with immunoprecipitation is trying to lower the number and type of background proteins that contaminate the washed immune complexes. Suggestions for decreasing background in IP. - [Read Troubleshoot Immunoprecipitation]

Articles (1-3 of 3)

Acid Guanidinium Thiocyanate RNA Isolation Phenol Chloroform Extraction

A single step RNA isolation protocol using Phenol Chloroform Extraction and Acid Guanidinium Thiocyanate. This RNA isolation method uses the fact that guanidinium thiocyanate can simultaneously lyse the cells and inactive cellular RNAses during the initial RNA isolation step allow a single step in the method.

In Vitro Translated Xenopus Mos Kinase Assay Protocol

In Vitro Translated Xenopus Mos Kinase Assay Protocol. In response to progesterone, immature Xenopus oocytes mature to eggs that can be fertilized. The Mos protein kinase is essential for oocyte maturation, most likely due to its ability to activate the MAP kinase cascade. This MAP kinase cascade eventually leads to the activation of Cdc2/cyclin B and entry into M phase. In this protocol, tagged Mos kinase is translated in vitro, immunopurified, and used in a kinase assay.

Vector Design Protocol for Transgenic Mice Generation

The protocol gives general considerations for the design of targeting vectors for transgenic mice. The protocol shares tips in the design of knock-out and knock-in vectors and some of their strategies for producing homologously recombined embryonic stem cells.

[1-3]