Protein Microarray Chips - An Introduction. Introduction, Types of Protein Chips, Attachment, Protein and Antibody Chip Production, Applications of Protein Chips, Detection methods, and Future Directions.
Molecularstation. - [Read Protein and Antibody Microarray Chips - An Introduction]
In this protocol, cells transfected with a luciferase reporter plasmid are lysed in a detergent-containing buffer. Luciferase in the extract catalyzes an oxidation reaction in which D-luciferin is converted to oxyluciferin, with production of light at 556 nm that can be quantified in a luminometer. - [Read Assay for Luciferase in Extracts of Mammalian Cells Protocol]
This protocol describes the isolation of fibroblasts from mouse embryos. Mouse embryonic fibroblast (MEF) cells are used as a feeder layer for the culture of mouse embryonic stem (ES) cells to help maintain them as pluripotent stem cells. The inhibition of ES-cell differentiation provided by the MEF feeders appears to be due to their production of leukemia inhibitory factor (LIF). - [Read Isolation and Freezing of Primary Mouse Embryonic Fibroblasts (MEF) For Feeder Plates]
Protocol for the expression of cloned genes in E. coli using IPTG-inducible promoters. Protocol describes how (1) to clone cloned sequences encoding open reading frames in plasmids carrying IPTG-inducible promoters, (2) to optimize expression of target proteins in transformants carrying these recombinants, and (3) to scale-up production of foreign proteins. - [Read Expression of Cloned Genes in E. coli Using IPTG-inducible Promoters Protocol]
Protocol for polyclonal antibody production. Very useful for rapid and simple generation of antibodies for western blots, ELISA assays, and immunoprecipitation. Includes: Rabbit Immunization; Initial Preparation; Pre-bleed; Antigen Injection; Monitoring of Titer; Purification of Antibodies. - [Read Polyclonal Antibody Production Protocol]
Antibody Array Production and Data Analysis. Probing Antigen Arrays and washing/ Blocking. Great Protocols and information. Brian Kidd. Stanford. - [Read Antibody Array Production and Analysis]
Protocol describes typical methods that are used to propagate and purify AAV vectors for experiments both in vitro and in vivo. Includes: Principles of the Triple Plasmid Transfection System; Plasmids; Transfection and Extraction of Virus; Purification of the AAV vector. - [Read A Protocol for AAV Vector Production and Purification]
Reference: Michael P. Matise, Wotjek Auerbach and Alexandra L. Joyner (2000). Gene targeting: a practical approach. Protocol excerpted from Chapter 3, Production of targeted embryonic stem cell clones. Alexandra L. Joyner (ed.), 2nd edition, Oxford Unive - [Read Embryonic Stem Cell growth media Requirements - Taconic Transgenics]
Protocol for steps in hybridoma production. Includes: SIMPLE METHOD: BY FROM GIBCO the HAT and HT concentrate; INSTRUMENTS FOR SPLENECTOMY OF ONE MICE; PREPARATION FOR CELL FUSION; CELL FUSION; SCREENING THE CLONES; RECLONING; ASCITIC FLUID PRODUCTION; PREPARATION OF PERITONEAL MACROPHAGES. - [Read Steps in Hybridoma Production Protocol]
Protocol is used to propagate and purify AAV vectors for experiments both in vitro and in vivo. Includes: Principles of the Triple Plasmid Transfection System; Plasmids; Transfection and Extraction of Virus; Purification of the AAV vector. - [Read AAV Vector Production and Purification Protocol]
Timeline for Transgenic Mouse Analysis and production. University of Michigan Transgenic Animal Model Core. Transgenic Animal Web. - [Read Timeline for Transgenic Mouse Analysis]
Production of T Cell Clones Protocol- https://catalog.invitrogen.com/index.cfm?fuseaction=iProtocol.unitSectionTree&treeNodeID=9E66347791E1CAD019BE41F3277340CA&objectid=6674BCB8F6BCF7B8032EEBAF7A3D08BD
Provides methods for the derivation of specific types of T cell clones: preparation and maintenance of alloreactive murine helper T (TH) lymphocyte and cytotoxic T lymphocyte (CTL) clones using the limiting dilution technique and derivation of TH clones reactive with soluble protein antigens including a method for the selection of either TH1 or TH2 lymphocyte subsets. - [Read Production of T Cell Clones Protocol]
For both biological and economical reasons, it is important to eliminate mycoplasmas from cell cultures being used for basic research, diagnosis, and biotechnological production. The most commonly used method for elimination, inactivation, or suppression of mycoplasmas in cell cultures is treatment with antibiotics. In general, antibiotic therapies do not result in long-lasting, successful elimination. Also, the cytotoxic properties of antibiotics can cause undesirable side effects on cells. - [Read Mycoplasma Elimination Reagent Protocol]
Assays for B Lymphocyte Function Protocols- https://catalog.invitrogen.com/index.cfm?fuseaction=iProtocol.unitSectionTree&treeNodeID=9E66340AD6485AA0FEB8F538B2FD389E&objectid=6674A538AC9B642C5CD51556EF7B3D26
Describes the antigenic stimulation of in vitro antibody production by B cells and the subsequent measurement of secreted antibodies. The first protocol is a generalized system for inducing in vitro antibody production and can accommodate various types of antigens under study. Secreted antibodies can then be measured with an enzyme-linked immunosorbent assay (ELISA) or other soluble-antibody detection systems. - [Read Assays for B Lymphocyte Function Protocols]
Protocol for the expression of cloned genes in E. coli using the bacteriophage T7 promoter. Protocol describes how (1) to clone cloned sequences encoding open reading frames in plasmids carrying bacteriophage T7 promoters, (2) to optimize expression of target proteins in transformants carrying these recombinants, and (3) to scale-up production of foreign proteins. - [Read Expression of Cloned Genes in E. coli Using the Bacteriophage T7 Promoter 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.
This Microarray Protocol Preparation of Fluorescent DNA Probes from Human mRNA protocol describes the production of probes labeled with the fluorescent dyes, Cy3 and Cy5, following the synthesis of cDNA from human mRNA and the hybridization of the probes to DNA microarrays.