allowed Protocols

Category: Cell Organelle Protocols: Subcellular Fractionation Protocols

Protocol describes systems in which the membranes are allowed to float through a density gradient from a dense load zone. This is an ideal format. - [Read A Self-generated Gradient to Discriminate a Cytosolic or Membrane Location of Large Protein Complex]

Category: Stem Cell Protocols: Stem Cell Culture Protocols

In vitro differentiation of ES cells occurs when the cells are allowed to aggregate in suspension culture in the absence of mouse embryonic fibroblast (MEF) feeders and leukemia inhibitory factor (LIF). Hanging drops provide a uniform aggregate size, which is then expanded by continued growth in suspension culture. The embryoid bodies are then plated and allowed to differentiate further in culture. - [Read Differentiation of Embryonic Stem (ES) Cells Using the Hanging Drop Method]

Category: Mass Spectrometry Protocols

Dried droplet Sample Preparation for MALDI.The discovery of this method allowed the application of laser desorption to proteins [2]. Drying a droplet of a protein/matrix solution remains the favorite method of most MALDI practitioners. Protocol for dried - [Read Dried droplet Sample Preparation for MALDI]

Category: Microscopy Protocols: Electron Microscopy Protocols

In recent years, the increased sensitivity of electron detectors and the availability of low-vacuum or variable-pressure systems have allowed imaging of fresh tissue samples without the need for fixation, drying, and coating. This obviously saves a lot of time, although the image quality may not be as good as that obtained from fixed samples. However, for most applications that tend to be at a relatively low magnification, the quality can be as good as that obtained from fixed samples. - [Read Imaging of Fresh Arabidopsis Tissues in the Scanning Electron Microscope]

Category: Plant Biology Protocols

In recent years, the increased sensitivity of electron detectors and the availability of low-vacuum or variable-pressure systems have allowed imaging of fresh tissue samples without the need for fixation, drying, and coating. This obviously saves a lot of time, although the image quality may not be as good as that obtained from fixed samples. However, for most applications that tend to be at a relatively low magnification, the quality can be as good as that obtained from fixed samples. - [Read Imaging of Fresh Arabidopsis Tissues in the Scanning Electron Microscope Protocol]

Category: Microscopy Protocols: Optical Microscopy Protocols

Diffraction-limited optical microscopy requires that the spatial resolution of an image is limited by the wavelength of the incident light & by the numerical apertures of the condenser & objective lens systems.The development of near-field scanning optical microscopy (scanning near-field optical microscopy) has allowed for a imaging technique that retains the various contrast mechanisms afforded by optical microscopy methods while attaining spatial resolution beyond the optical diffraction limit - [Read Near-Field Scanning Optical Microscopy]

Category: Stem Cell Protocols

This protocol describes passage of ES cells. They should be split at 1:3 to 1:7 every 2-3 days depending on their growth rate when they reach 70% confluency. They should never be allowed to grow past 90% confluency, but rather they should form tightly packed colonies not touching each other. - [Read Passage of Embryonic Stem (ES) Cells Protocol]

Category: Mouse Protocols: Mouse Cell Culture Protocols

The liver of a rat is cannulated and perfused in situ with buffer, following which it is excised and perfused in a closed system with a collagenase solution. After a period of time the liver begins to break up, at which point it is transferred to a measuring cylinder and culture medium is added. It is then gently agitated to cause the release of cells which are subsequently filtered and allowed to settle out. The parenchymal and non-parenchymal cells form two distinct layers which can be separat - [Read The Isolation and Culture of Rat Hepatic Cells Protocol]

Category: Epigenetics and Methylation Protocols: 5-Aza-dC Treatment AZA Procedure

"Cells were seeded at a density of 1X106 cells/100-mm dish with 10% FBS and allowed to attach over a 24-h period. 5-Aza-dC (Sigma) was then added to a final concentration of 0.2–1
M, and the cells were allowed to grow for 5 days. The medium with or w - [Read Treatment of Cells with 5-aza-dC. protocol PDF]