To 2mls of mid-log culture or 1ml of freshly saturated culture add 1ml(or an equal volume) of glycerol solution, mix gently, then freeze rapidly in liquid nitrogen or on dry ice. Store at -20°C and - 70°C. - [Read Bacterial Glycerol Stocks Protocol]
Freezing gealthy cells (> 95% viable) the protocol obtains a culture 80-90% viable 24 hours after thawing and growing on test vial from step 9. Antisense Research Group - [Read Cell Freezing using Liquid Nitrogen N2]
Culture of Human Prostatic Carcinoma Cell Lines,
Growing and splitting the cells, Preparation of frozen stocks in liquid nitrogen, How to bring frozen cells back into culture, Concentrations of antibiotics for the selection of stable transfectants. LNCa - [Read Culture of Human Prostatic Carcinoma Cell Lines]
This protocol has been used successfully to 15N or 13C/15N label our proteins using our pET1120/BL21(DE3) expression system: Preparing M9 minimal media begins with preparing a 5x stock solution of M9 salts. Generally, M9 salts contain a nitrogen source in the form of NH4Cl. Since we want to add a labeled nitrogen source, our 5x salts are prepared minus NH4Cl. Standard 5 X M9 Minimal Media salts minus nitrogen source For 1L 5xM9 salts: - [Read Expression Protocol in M9 Minimal Media via T7 Promoter]
Freezing and Thawing Eukaryotic Cells in liquid nitrogen, Freezing Cell Lysates for Western Blots, Thawing Cells from Liquid Nitrogen. Dr. Bart Frank Oklahoma. - [Read Freezing and Thawing Eukaryotic Cells]
Freezing and Thawing of Mammalian Cell Lines. For long term storage of myeloma cells, hybridoma cells, T cells, and other mammalian cell lines in liquid nitrogen, and restoring them in culture. Kitto Lab Texas. - [Read Freezing and Thawing of Mammalian Cell Lines]
Protocol for Protein Extraction Using Proteomics. Extraction of proteins from plant cells that are rich in compounds that interfere with the 2-Dimensional electrophoretic separation methods such as salts, organic acids, phenolics, pigments, terpenes, among others. A common protocol used in our lab for extraction proteins from plant tissues consists in the homogenization of mortar-grounded material in liquid nitrogen with an extraction buffer. - [Read Protocol for Protein Extraction Using Proteomics]
Tubulin is polymerized into microtubules by incubating tubulin at 37°C with GTP. A nucleation seed is added when the purpose is to assay microtubule elongation. Tubulin can also be polymerized for the purposes of recycling the tubulin or labeling the microtubules with fluorescently labeled tubulin. Based on the protocol by Timothy Mitchison of Harvard University.
Histone H1 Kinase Activity Assay Protocol. This protocol describes assaying kinase activity of a putative kinase using Histone H1 as the substrate. Histone H1 is the canonical kinase substrate in this type of assay. Phosphorylation of Histone H1 is assessed by SDS-Polyacrylamide gel electrophoresis followed by autoradiography.