transient Protocols

Category: Cell Biology and Cell Culture: Apoptosis Protocols: Apoptosis Analysis

An Integrative Procedure for Apoptosis Identification and Measurement. Yingyu Cui Lab/Group: National Key Laboratory of Protein Engineering and Plant Genetic Engineering, College of Life Sciences. I have uploaded an integrative procedure through which a relatively satisfactory result can be obtained following a single stage of cell culture and transient cell treatment, then detection with different instruments. This shortens experiment time. - [Read An Integrative Procedure For Apoptosis Identification And Measurement]

Category: Cell Biology and Cell Culture: Flow Cytometry Protocols: Apoptosis Cytometry Protocols

An integrative procedure through which a relatively satisfactory result can be obtained following a single stage of cell culture and transient cell treatment, then detection with different instruments. This shortens experiment time. - [Read Apoptosis Identification and Measurement Protocol]

Category: Transfection Protocols: Calcium Phosphate Transfection Protocols

This calcium phosphate transfection method works best in cell lines that are 1) highly transformed and 2) adherent (Hela, U2OS, SAOS2, AdAH, NPC-KT and obtain from 20% to 100% transfection efficiency depending on the cell line). Works well for transient experiments but precautions should be used in the design and interpretation of experiments based on the discussion below. Also works very well for generating stable cell lines. This method is quite sensitive to the amount of input plasmid. - [Read Calcium Phosphate Transfection Method]

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: Cell Biology and Cell Culture: Cell Transfection Protocol

Lipofectamine transient tranfection of mammalian cells in IMDM cell media. Julie B. Wolf, UMBC. - [Read Procedure for Transfection of Mammalian Cells Using Lipofectamine]

Category: Yeast Protocols: Yeast Genetics Protocols: Yeast Replication Protocols

Protocol for replication timing using transient hemimethylation. Includes: Strain construction; Protocol for methylation timing; Quantitation. - [Read Replication Timing Using Transient Hemimethylation Protocol]

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

GUS is used as a tag to address nuclear localization whereas GFP is more versatile. GFP is detectable directly in living cells, GUS is only detected indirectly by staining of fixed tissue which may lead to artifacts or may obscure problems with protein solubility. In this protocol, protein localization is routinely assayed after particle-mediated transient transformation of onion epidermal cells. With this method it can be determined rapidly whether a given fusion protein is active and.... - [Read Subcellular Localization of GUS- and GFP-Tagged Proteins in Onion Epidermal Cells]

Category: Transfection Protocols

DEAE-dextran is generally used to obtain a burst of transient expression of cloned genes after transfection of mammalian cells. Many variants of the technique have been described, all of which seek to maximize the uptake of DNA and to minimize the cytotoxic effects of DEAE-dextran. In this protocol cells are exposed briefly to a high concentration of DEAE-dextran-DNA and then to chloroquine diphosphate, which is a facilitator of transfection. - [Read Transfection Mediated by DEAE-Dextran: High-efficiency Method Protocol]

Category: Plant Biology Protocols: Plant Genetics Protocols

The study of transient gene expression provides a useful complement to the study of stably transformed plants. Transient assays offer a quick method of testing the effects of genes, using either phenotypic, molecular, or biochemical readouts. Transient assays based on Agrobacterium-mediated transformation of leaf explants have been described for other plant species, but it is not known how well these assays work in Arabidopsis. - [Read Transient Expression in Protoplasts]

Category: Transfection Protocols: Transient Transfection Protocols

Transient transfection into mammalian cells is a convenient way to over express and obtain protein expression. Protocol includes: Culture conditions; Transfection of experimental cells; Preparation of Mammalian Cell Lysate for Luciferase assay. - [Read Transient Transfection and Luciferase assay]

Category: Reporter Gene Assays: Luciferase Assay Protocols

Transient transfection into mammalian cells is a convenient way to over express and obtain protein expression. Protocol includes: Culture conditions; Transfection of experimental cells; Preparation of Mammalian Cell Lysate for Luciferase assay. - [Read Transient Transfection and Luciferase Assay Protocol]

Category: Transfection Protocols: Transient Transfection Protocols

Protocol for transient transfection and spin infection. - [Read Transient Transfection and Spin Infection Protocol]

Category: Cell Biology and Cell Culture: Cell Transfection Protocol

Transient transfection into 293T cells Protocol. transfection, calcium phosphate, overexpression, protein production. Jun Takagi - [Read Transient transfection into 293T cells]

Category: Transfection Protocols: Transient Transfection Protocols

Transient transfection into 293T cells is a convenient way to overexpress and obtain both cellular and extracellular (secreted or membrane) proteins. 293 is a human renal epithelial cell line which is transformed by adenovirus E1A gene product. 293T is a derivative which also express SV40 large T antigen, allowing episomal replication of plasmids containing the SV40 origin and early promoter region. They (both) have the unusual property of being highly transfectable. - [Read Transient Transfection Into 293T Cells Protocol]

Category: Transfection Protocols: Transient Transfection Protocols

Protocol for the transient transfection of COS cells. - [Read Transient Transfection of COS Cells Protocol]

Category: Cell Biology and Cell Culture: Cell Transfection Protocol

Protocol optimized for Cos-1 cells. Successful transfection of each cell type requires optimization of the basic protocol. LIPOFECTAMINE method of transient transfection. Upstate. - [Read Transient Transfection of Cos-1 Cells]

Category: Transfection Protocols: Transient Transfection Protocols

Transient transfection protocol for the ecotropic/amphotropic producer line including titering and concentration of viral stocks. - [Read Transient Transfection Protocol: Viral Production]

Category: Transfection Protocols: Transient Transfection Protocols

This transient transfection protocol for the ecotropic/amphotropic producer line includes instructions for titering and concentration of viral stocks. This procedure gives consistently higher transfection efficiency. - [Read Transient Transfection: Viral Production]