ES Cell Culture Protocols - EXCELLENT GUIDE. Harvesting and DNA Preparation, Colony Picking, Electroporation, ES Cell Culture using ESGRO® and Feeder Cell Layer. ESGRO. - [Read ES Cell Culture Protocols - ESGRO]
This protocol describes a method for recombining and culturing germ layer fragments. It is useful for testing the inductive properties of fragments from wild-type and mutant mouse embryos. - [Read Germ Layer Explant Recombination Culture Protocol]
Protocol describes a method for recombining and culturing germ layer fragments. It is useful for testing the inductive properties of fragments from wild-type and mutant mouse embryos. - [Read Germ Layer Explant Recombination Culture Protocol]
Immunostaining thin layer chromatograms TLC is a very sensitive detection technique of functionally active carbohydrate ligands of protein receptors. Carbohydrate structures are detected in glycolipids from complex mixtures of molecules extracted from the relevant target tissue. Proteins analyzed can be antibodies, chimeric Ig proteins, selectins, lectins, toxins, and other carbohydrate binding proteins. John L. Magnani~GlycoTech Corporation, Rockville, Maryland - [Read Immunostaining Thin Layer Chromatograms Of Glycolipids]
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]
In this protocol, extracts prepared from cells transfected with a chloramphenicol acetyltransferase (CAT) reporter plasmid are incubated with radiolabeled chloramphenicol. The acetylated products generated by the action of CAT are separated from the unmodified drug by thin-layer chromatography and quantitated by scraping the spots from the thin-layer plates and counting them by scintillation spectroscopy. - [Read Measurement of CAT in Extracts of Mammalian Cells Using Thin-layer Chromatography]
Gliotoxin is a metabolite of Aspergillus fumigatus that exhibits immunosuppressive activity against certain cells of the immune system. Secretion of gliotoxin during infection has been suggested as being a factor in the pathogenesis of aspergillosis. Gliotoxin secretion can be assayed in a number of ways by thin layer chromatography (TLC) high performance liquid chromatography (HPLC) or bioassay using the effect of gliotoxin on human cells1. - [Read Method for Assaying Gliotoxin Production in Aspergillus fumigatus Protocol]
Details a placenta specific gene manipulation by transducing blastocysts with lentiviral vectors1. After a removal of zona pellucida which functions as a physical barrier, trophoblast cells lying outermost layer of blastocyst were transduced from outside with high-titer lentiviral vectors. As most placental cells descend from trophoblast cells while fetus originated from inner cell mass, transgene expression can be observed in trophoblast cells from preimplantation stages and in placenta... - [Read Placenta Specific Gene Manipulation by Transducing Zona-Free Blastocyst using Lentiviral Vector]
This method of sample preparation produces a uniform layer of very small crystals on the mass spectrometer's sample stage that are mechanically well adhered to the substrate. It is a variant of the dried droplet method. PROWL - [Read Polycrystalline thin films Sample Preparation Protocol MALDI]
This protocol describes the culture of embryonic stem (ES) cells using mitotically inactivating primary mouse embryonic fibroblast (MEF) cells as a feeder layer (preparation described in Preparation of Mouse Embryonic Fibroblast (MEF) Feeder Plates). The ES culture medium is supplemented with recombinant leukemia inhibitory factor (LIF) to help maintain the cells as pluripotent stem cells. This protocol has been optimized for the ES-D3 cell line. - [Read Propagation of Pluripotent Mouse Embryonic Stem (ES) Cells Protocol]
This protocol describes the culture of embryonic stem (ES) cells using mitotically inactivating primary mouse embryonic fibroblast (MEF) cells as a feeder layer (preparation described in Preparation of Mouse Embryonic Fibroblast (MEF) Feeder Plates). The ES culture medium is supplemented with recombinant leukemia inhibitory factor (LIF) to help maintain the cells as pluripotent stem cells. This protocol has been optimized for the ES-D3 cell line. - [Read Propagation of Pluripotent Mouse Embryonic Stem (ES) Cells Protocol]
This protocol a protocol on how to generate transfected embryonic stem (ES) cell clones. The previous protocol in this series is the Protocol for Electroporation of ES cells. The next protocol in the series is the Protocol on Disaggregation, Expansion, and Freezing of Transfected ES Clones.
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.