transfer Protocols

Category: Primary Cell Isolation and Culture Protocols: Laser Capture Microdissection Protocols

Protocol for 2-D polyacrylamide gel electrophoresis. Includes: 50 ml IEF Lysis Buffer; 10X Electrophoresis Running Buffer (10 L); 30% Acrylamide Stock (1 liter); Separating Acrylamide Gel; 50 ml Equilibration Buffer I; 50 ml Equilibration Buffer II; Transfer Buffer; Sample Preparation; Tissue Processing; Reswelling; Prepare Gradient Acrylamide Gel (9-18%); Transfer and Sequencing of Proteins. - [Read 2-D Polyacrylamide Gel Electrophoresis Protocol]

Category: Yeast Protocols: Yeast Nucleic Acid Protocols: Yeast PCR Protocols

Protocol for 96-well confirmation Yeast PCR. Includes: Clonal purification; Generate a master plate (96-well format); Making a frozen backup stock; Confirmation PCR for one Row; ORF Specific Confirmation PCR --> "A-B" primers (upstream junction); Transfer template DNA to multiwell PCR plate; Prepare and dispense master mix for A-B PCR. - [Read 96-Well Confirmation Yeast PCR Protocol]

Category: Plant Biology Protocols: Plant Genetics Protocols

Protocol describing Agrobacterium mediated gene transfer via hypocotyls. - [Read Agrobacterium-Mediated Gene Transfer via Hypocotyls Protocol]

Category: Mouse Protocols: Mouse Embryology Protocols

Protocol for blastocyst embryo transfer into pseudopregnant recipient female mouse. - [Read Blastocyst Embryo Transfer into Pseudopregnant Mouse Protocol]

Category: Cell Biology and Cell Culture: ES Stem Cell Biology and Culture

Blastocyst Transfer. Bowtell Lab. - [Read Blastocyst Transfer.]

Category: Plant Biology Protocols: Plant Proteins, Peptides and Antigens

BN-PAGE has become the method of choice for the investigation of the respiratory protein complexes of the electron transfer chains of a range of organisms. It allows the separation in two dimensions of extremely hydrophobic protein sets for analysis and also provides information on their native interactions. In this review we discuss the capabilities of BN-PAGE in proteomics and the wider investigation of protein:protein interactions with a focus on its use and potential in plant science. - [Read Blue-Native PAGE in Plants: A Tool in Analysis of Protein-Protein Interactions]

Category: Developmental Biology: Utilizing Model Organisms Protocols

In the experiment there are three players A, B, and C. Player A gets an initial endowment of 10, player B gets 0 and player C gets 5 points. First, player A decides how many of his 10 points to transfer to B. Then, player C observes the decisions of A and gets the possibility to leave the payoffs unaffected or to punish A by deducting 3 (or 6) points from A at a cost of 1 point (or 2 points) for C.... - [Read Experiments on Parochial Altruism in Humans; Procedures and Instructions]

Category: Yeast Protocols: Yeast Genetics Protocols: YACs Protocols

Protocol describes methods for isolation of DNA from a strain of S. cerevisiae carrying a recombinant YAC. Because the linear YAC DNAs are sensitive to shearing forces, pipettes with wide-bore tips should be used to transfer DNAs. The method is suitable for preparing DNA that will be used for agarose gel electrophoresis, Southern blotting, subcloning, genomic library construction, PCR, or other methods that do not require intact high-molecular-weight DNA. - [Read Growth of S. cerevisiae and Preparation of DNA Protocol]

Category: Western Blot Protocols

Protocol for high throughput western blotting for antibody quality control. Includes: SDS-PAGE; TRANSFER; IMMUNOSTAINING; DETECTION. - [Read High Throughput Western Blotting Protocol for Antibody Quality Control]

Category: Primary Cell Isolation and Culture Protocols: Laser Capture Microdissection Protocols

Protocol for Immunoblot. Includes: Staining and Laser Capture Microdissection; Protein Separation by Polyacrylamide Gel Electrophoresis; Electrophoretic Transfer To a Membrane (Nylon, PVDF or Nitrocellulose); Primary and Secondary Antibody Incubations; Visualization. - [Read Immunoblot Protocol]

Category: Western Blot Protocols: General Western Blot Methods Protocols

The transfer of proteins from a Tris/glycine SDS-polyacrylamide gel to a membrane using a semi-dry method is achieved by placing the gel next to a piece of nitrocellulose filter.  This sandwich is placed directly between two plate electrodes, and the proteins are then transferred from the gel onto the filter. - [Read Immunoblotting: Semi-Dry Electrophoretic Transfer of Proteins from Gels to Membranes Protocol]

Category: Western Blot Protocols: General Western Blot Methods Protocols

Transfer of proteins from a Tris/glycine SDS-polyacrylamide gel to a membrane using the submerged method is achieved by placing the gel next to a piece of nitrocellulose filter, submerging this sandwich in a large volume of transfer buffer in a transfer tank, and running current from one side of the transfer tank to another.  The proteins are then eluted by transferring them from the gel onto the filter. - [Read Immunoblotting: Submerged Electrophoretic Transfer of Proteins from Gels to Membranes Protocol]

Category: Immunology: T Cell Protocols

Describes methods for labeling high or low numbers of lymphocytes with CSFE. Protocols are provided to use CSFE-labeled cells in cell transfer studies or as cells to be cultured in vitro. Detailed guidelines for positioning of CSFE-labeled lymphocytes in lymphoid organs or other tissues are included for those wishing to use this approach to study lymphocyte migration. - [Read Intracellular Fluorescent Dye CFSE to Monitor Lymphocyte Migration and Proliferation]

Category: Nucleic Acid Purification Protocols: DNA Isolation Protocols: DNA Isolation from Mammalian Cells Protocols

Method of choice when large amounts of mammalian DNA are required, for example, for Southern blotting (Rapid Isolation of Mammalian DNA, Rapid Isolation of Yeast DNA, Southern Blotting: Capillary Transfer of DNA to Membranes) or for construction of genomic libraries in bacteriophage {lambda} vectors.  Approximately 200 µg of mammalian DNA, 100-150 kb in length, is obtained from 5 x 107 cultured aneuploid mammalian cells (e.g., HeLa cells). - [Read Isolation of High-molecular-weight DNA from Mammalian Cells Using Proteinase K and Phenol Protocol]

Category: Molecular Model Organisms: Mouse Animal Protocols: Mouse Genotyping Protocols

Contains the NHLBI Division of Intramural Research, including a list of labs, and the Office of Technology Transfer and Development. - [Read Laboratory of Developmental Biology, Division of Intramural Research, NHLBI, NIH]

Category: Immunology

A method for ameliorating autoimmune disease by passive transfer of IVIg-primed leukocytes. Developed a method for ‘priming’ specific leukocyte populations with IVIg to study their potential role in ameliorating ITP upon passive transfer, which we describe herein. Using this technique, found that splenic CD11c+ dendritic cells reacted with IVIg, or several IVIg mimetic regimes, were primed to ameliorate ITP upon transfer to thrombocytopenic mice. - [Read Method for Ameliorating Autoimmune Disease by Passive Transfer of IVIg-Primed Leukocytes]

Category: RNA Protocols: cDNA Libraries Library

Purpose: To transfer DNA from bacterial colonies to nylon (or nitrocellulose) membrane so that a library can be screened by hybridization. Jim Howe Donis Keller Lab - [Read Method: Transfer of DNA from Bacterial Colonies to Nylon Membrane or Nitrocellulose]

Category: Mouse Protocols: Mouse Reproductive System Surgery Protocols

Protocol describes a method for Caesarean section and fostering. Caesarean section is required if the recipient of an embryo transfer or any pregnant mouse has not given birth by the delivery time normal for the particular strain. - [Read Mouse Caesarean Section and Fostering Protocol]

Category: Mouse Protocols: Mouse Embryology Protocols

1-cell embryo transfer into pseudopregnant recipient female mouse. - [Read Mouse Embryo Transfer into Pseudopregnant Recipient Protocol]

Category: Fungi Protocols: Neurospora Protocols

Northern blot protocol for the detection of RNA in Neurospora. Includes: Extract RNA from tissue powder; Electropheresis and transfer RNA; Preparing the membrane for probing and preparation of a riboprob.; Hybridization. - [Read Northern blot Protocol for the Detection of RNA in Neurospora]

Category: Immunology: T Cell Protocols

Protocol for nucleofection and adoptive transfer of primary mouse T lymphocytes. - [Read Nucleofection and Adoptive Transfer of Primary Mouse T Lymphocytes Protocol]

Category: Transfection Protocols: Transient Transfection Protocols

Transient transfection of genes into naïve mouse T cells using nucleofection using a modified electroporation technique. This protocol offers the possibility for rapid functional in vivo studies of targeted genes in primary mouse T cells. - [Read Nucleofection and Adoptive Transfer of Primary Mouse T lymphocytes using Transient Transfection]

Category: Protein Protocols: Western Blot Protocol: Troubleshooting Western Blots

Optimizing Western Blots: Pre-Transfer Invitrogen - [Read Optimizing Western Blots: Pre-Transfer Invitrogen]

Category: Mouse Protocols: Mouse Reproductive System Surgery Protocols

Protocol describes oviduct transfer of mouse embryos. It is based on the Whittingham method, which applied a well described procedure for the rat to the mouse. It is best to practice this procedure first on a cadaver and then on an anesthetized 0.5-dpc pseudopregnant mouse using a dye solution or blue Affigel beads rather than embryos to gain experience in finding the opening of the oviduct (infundibulum). - [Read Oviduct Transfer Protocol]

Category: Gene Delivery Protocols: DNA Nanoparticles Protocols

This protocol describes the preparation of polyethylenimine (PEI)/DNA nanoparticles for targeted gene delivery. This delivery strategy improves the efficiency of gene transfer by enhancing the entry of gene vectors into the desired cells and reducing uptake by nontarget cells. We describe here methods for the conjugation of targeting peptides to PEIs, formation of DNA complexes using the conjugated PEIs or nonconjugated PEIs together with targeting peptides, and cell transfection. - [Read PEI Nanoparticles for Targeted Gene Delivery Protocol]