Collagenase perfusion of rat liver yields a hepatocyte suspension which may be exposed to test compounds in order to assess their effects on cell viability and enzyme leakage. - [Read Isolation of Rat Hepatocytes Protocol]
Describes the isolation of resting B lymphocytes (B cells) from mouse spleens by using negative selection with anti-CD43 and anti-Mac-1/CD11b monoclonal antibodies coupled to magnetic microbeads. This strategy depletes non-B cells from a mixed population of splenocytes and relies on the fact that most mature leukocytes, with the exception of resting splenic B cells, express CD43. - [Read Isolation of Resting B Lymphocytes from One or More Groups of Four Mouse Spleens Protocol]
Protocol describes the isolation of resting B lymphocytes (B cells) from mouse spleens by using negative selection with anti-CD43 and anti-Mac-1/CD11b monoclonal antibodies coupled to magnetic microbeads. This strategy depletes non-B cells from a mixed population of splenocytes and relies on the fact that most mature leukocytes, with the exception of resting splenic B cells, express CD43. - [Read Isolation of Resting B Lymphocytes from Sixteen Mouse Spleens Protocol]
Describes the isolation of resting B lymphocytes (B cells) from mouse spleens by using negative selection with anti-CD43 and anti-Mac-1/CD11b monoclonal antibodies coupled to magnetic microbeads. This strategy depletes non-B cells from a mixed population of splenocytes and relies on the fact that most mature leukocytes, with the exception of resting splenic B cells, express CD43. - [Read Isolation of Resting B Lymphocytes from Sixteen Mouse Spleens Protocol]
This procedure describes the isolation of resting B lymphocytes (B cells) from mouse spleens by using negative selection with anti-CD43 and anti-Mac-1/CD11b monoclonal antibodies coupled to magnetic microbeads. This strategy depletes non-B cells from a mixed population of splenocytes and relies on the fact that most mature leukocytes, with the exception of resting splenic B cells, express CD43. - [Read Isolation of Resting B Lymphocytes Protocol]
Retroelements and their derivatives are a ubiquitous and abundant component of plant genomes. Major classes of retroelements include the Pseudoviridae (Ty1-copia ), the Metaviridae (Ty3 -gypsy) and the Retroposineae LINE (non-LTR) groups. All reverse transcribing elements can be included in a universal classification. Includes: Pseudoviridae (Ty1-copia) Degenerate Primers; Metaviridae (Ty3-gypsy) Element Degenerate Primers; LINE Element Degenerate Primers; PCR Programmes. - [Read Isolation of Retroelement from Plant Genomic DNA]
Protocol for isolation of T cells from synovial membranes. Includes: Initial Cell Isolation; Nonadherent Cell Isolation; Adherent Cell Isolation; Neuraminidase-Treated Sheep Red Blood Cells; Erythrocyte Lysing Solution; Balance Salt Solution (BSS). - [Read Isolation of T Cells from Synovial Membranes Protocol]
A discontinuous gradient of iodixanol is used in which the crude mitochondrial fraction is layered beneath the preformed
gradient. In this protocol the osmolality of the gradient is approx 600 mOsm and the crude mitochondrial fraction is loaded in 40% iodixanol rather than 50% iodixanol. - [Read Isolation of Yeast Mitochondria in aPre-Formed Iodixanol Gradient]
Protocol describes the use of agarose plugs for isolation of yeast artificial chromosome (YAC) DNA. The DNA can then be run on a pulsed-field gel and used for microinjection to produce transgenic mice. - [Read Large-Scale Preparation of Agarose Plugs of Yeast DNA Protocol]
Protocol describes a method for isolation and stimulation of lymphocytes. Leukocyte-rich plasma is collected from whole blood and then centrifuged through Ficoll-Hypaque. The collected cells are resuspended in growth medium containing various mitogens to stimulate growth. - [Read Lymphocyte Isolation and Culture Protocol]
Protocol is the first step in a three-step process for the preparation and enrichment of phosphopeptides using immobilized metal affinity chromatography (IMAC) for the identification of the phosphopeptides by LC-MS/MS. This procedure is used to prepare protein extracts from WEHI-231 cells. This preparation method provides total cellular protein samples that are free of contaminating nucleic acids. - [Read Lysis and Protein Extraction from WEHI-231 Cells with TriPure Isolation Reagent Protocol]
Microneedles attached to micromanipulators are used in the dissection of tetrads, isolation of zygotes from populations of mating haploid cells, and manipulation of individual cells. - [Read Making a Tetrad Dissection Needle Protocol]
Large scale double-stranded DNA isolation, Miniprep double-stranded DNA isolation, Large scale M13RF isolation, Single-stranded M13 DNA isolation using phenol. Minion Lab.Veterinary Medicine Iowa Univ. - [Read Methods for DNA isolation]
Large scale double-stranded DNA isolation, Midiprep double-stranded DNA isolation. Bruce A. Roe, Department of Chemistry and Biochemistry, The University of Oklahoma, Norman. - [Read Methods for DNA isolation - Protocol Book]
This protocol describes the isolation of microsomal Endoplasmic Reticulum (ER) membranes from yeast cells. Includes: Spheroplast Formation and Microsome Purification. - [Read Microsome Preparation from Yeast Cells Protocol]
A Single Stranded Plasmid DNA Isolation Protocol describing the production and isolation of single-stranded DNA (ssDNA) using bacteriophagemid-containing bacteria and helper phage. Infection of the host cells with helper phage allows for packaging of ssDNA into bacteriophage. The ssDNA can then be isolated from phage particles.
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.
The DNA Ligation protocol described here contains the steps required to join together using ligase enzyme both plasmid DNA and insert DNA fragments in order to create a new plasmid. This new ligated plasmid can be transformed after into competent bacteria to produce DNA for mini, midi or maxi-prep isolation.
This protocol describes the electroporation of the BMH 81-17 mut S strain that is recommended for tranformation of the site directed mutagenesis of dsDNA (See Protocol on Site-Directed Mutagenesis on Double Stranded DNA). BMH 81-17 mut S are a mismatch repair defective (mut S) Escherichia coli strain. The probability that the two mutations will cosegregate during the first round of DNA replication is increased in this strain.