293t Protocols

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Northern Blot Analysis of mRNA From Mammalian Polyribosomes Protocol - http://www.natureprotocols.com/2006/09/15/northern_blot_analysis_of_mrna.php

Category: Nucleic Acid Detection Protocols: Nucleic Acid Blotting Protocols

The protocol consists of a method for the generation of cytoplasmic extracts from mammalian cells (in this case, 293T cells) without the disruption of polyribosomes, the separation of ribosomal components and polyribosomes by sucrose gradient centrifugation, the isolation of mRNA from these fractions, and detection of mRNA by Northern blot analysis. - [Read Northern Blot Analysis of mRNA From Mammalian Polyribosomes Protocol]

Transfection/Retroviral Infection pCL-Eco+Gene of Interest into 293T Cells Infecting HC11 Cells - http://www.bcm.edu/rosenlab/protocols/retroviral.pdf

Category: Cloning Protocols: Vector Protocols

Protocol for the transfection/Retroviral Infection pCL-Eco+Gene of interest into 293T cells infecting HC11 cells. Includes: Split 293T cells for transfection; Transfect 293T cells; Split HC11 cells for infection; Infect target (HC11) cells. - [Read Transfection/Retroviral Infection pCL-Eco+Gene of Interest into 293T Cells Infecting HC11 Cells]

Transient transfection into 293T cells - http://www.cbrinstitute.org/labs/springer/protocols/jun_293T.html

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]

Transient Transfection Into 293T Cells Protocol - http://www.cbrinstitute.org/labs/springer/protocols/jun_293T.html

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]

Western Blot Analysis of Sub-Cellular Fractionated Samples Using the Odyssey Infrared Imaging System - http://www.natureprotocols.com/2006/09/15/western_blot_analysis_of_subce.php

Category: Cell Organelle Protocols: Subcellular Fractionation Protocols

This protocol describes nuclear and cytoplasmic fractionation of tissue culture cells and a method for Western blot detection of proteins using the Odyssey Infrared Imaging System. This protocol was used to detect expression of the "small" Tap protein in 293T, HeLa and COS cells. The Odyssey system has several advantages over the more widely used chemiluminescent detection methods. - [Read Western Blot Analysis of Sub-Cellular Fractionated Samples Using the Odyssey Infrared Imaging System]