Chromatin immunoprecipitation protocol to analyze histone modifications in Arabidopsis thaliana. Werner Aufsatz, Matzke Lab. Gregor Mendel Institute of Molecular Plant Biology, Austria. Includes Chromatin Crosslinking, Chromatin preparation, Pre-clearing and immuno precipitation (IP), Collection,washes and elution of immune complexes, Reverse crosslinking and DNA cleanup - [Read Chromatin immunoprecipitation protocol to analyze histone modifications in Arabidopsis thaliana]
In this protocol, the DNA-binding capacity of Wizard MagneSil particles is used to capture and release a consistent amount of DNA (100 ng) across a wide range of samples. At the end of the procedure, the DNA is eluted into 100 µl Elution Buffer to give a final concentration of 1 ng/µl, relieving the need for postpurification DNA quantitation. - [Read DNA IQ Isolation of Genomic DNA from Stains and Buccal Swabs Protocol]
Protocol for the isolation of infectious HIV-1. Includes: HIV-1 capture strategy; Protocol for the isolation of HIV-1 virions; Magnetic labeling; Magnetic separation; Elution option A for virion lysate; Elution option B for intact virions. - [Read Isolation of Infectious HIV-1 Protocol]
Protocol for the optimization of elution condition for dye-ligand affinity chromotography. Elution methods used in dye-ligand affinity chromatography may be either selective or nonselective in nature. Usually, selective elution methods are applied in combination with group-specific adsorbents, such as dye-ligand adsorbents, and nonselective elution methods are used in combination with highly specific adsorbents. - [Read Optimization of Elution Conditions for Dye-Ligand Affinity Chromatography Protocol]
Recombinant proteins, constructed in pGEX vectors, are fused to glutathione S-transferase (GST) and can be purified to near homogeneity by affinity chromatography on glutathione-agarose. Bound GST-fusion proteins are readily displaced from the column by elution with buffers containing free glutathione. - [Read Purification of Fusion Proteins by Affinity Chromatography on Glutathione Agarose Protocol]
Immobilized metal-ion affinity chromatography (IMAC) is suitable for the purification of proteins under denaturing conditions. Either guanidine-HCl or urea can be used, although guanidine-HCl is a stronger denaturant than urea. Proteins that have been adsorbed to the column in the presence of guanidine-binding buffer may be washed with urea-binding buffer and eluted with urea elution buffer. - [Read Purification of Histidine-Tagged Proteins under Denaturing Conditions Using IMAC Protocol]
Rapid Elution of DNA Agarose Gels Protocol. This method allows quick purification of DNA fragments from agarose gels for use in cloning and other reactions. Higher yields of purified DNA can be obtained from commercially available purification kits, however greater ligation efficiencies per given amount of DNA have been seen with the use of these described spin columns. Hahn Lab. - [Read Rapid Elution of DNA Agarose Gels Protocol]
Chromatography on oligo(dT) columns is the preferred method for large-scale purification (>25 µg) of poly(A)+ RNA extracted from mammalian cells. Typically, between 1% and 10% of the RNA applied to the oligo(dT) column is recovered as poly(A)+ RNA. Because the method can be frustratingly slow, it is not recommended for purification of poly(A)+ RNA from multiple samples. For this purpose, batch elution (Selection of Poly(A)+ RNA by Batch Chromatography) is the better choice. - [Read Selection of Poly(A)+ RNA by Oligo(dT)-Cellulose Chromatography - Subscription Required]
Ultimate His-UB Assay for Mammalian Cells. Tansey Lab Protocols. William P. Tansey. PREPARATION OF NI-NTA-AGAROSE, HARVESTING THE TRANSFECTION., ELUTION OF HIS-TAGGED PROTEINS. - [Read Ultimate His-UB Assay for Mammalian Cells]
A protocol for the selection of Phage Antibodies using Immobilized Antigen. This method describes the selection of antibodies from bacteriophage antibody libraries that recognize a specific antigen. The phage display library of antibody-displaying phage particles is exposed to antigen attached to a solid substrate (Nunc Immuno™ tubes). The phage particles with affinity for antigen bind to the immobilized antigen and are selected from the library of phage expressing antibodies.