enter Protocols

Category: Reporter Gene Assays: B-Galactosidase Assay Protocols

In the method described here, the cells are permeabilized to allow the substrate to enter the cells and the activity is normalized to the number of cells assayed. - [Read Assay of ß-Galactosidase in Yeast: Permeabilized Cell Assay]

Category: Yeast Protocols: Yeast Genetics Protocols: B-Galactosidase Assay Protocols

In this protocol, the cells are permeabilized to allow the substrate to enter the cells and the activity is normalized to the number of cells assayed. - [Read Assay of ß-Galactosidase in Yeast: Permeabilized Cell Assay Protocol]

Category: Cell Biology and Cell Culture: Flow Cytometry Protocols

FIXATION and DNA Staining for Cell Cycle Analysis Protocol. This method of DNA staining utilizes ethanol to fix the cells and permeabilize the membrane, which allows the dye (Propidium Iodide) to enter the cells.  Propidium Iodide (PI) is a DNA-binding fluorochrome that intercalates in the double-helix.  Ribonuclease-A is used to eliminate the staining of double-stranded RNA. - [Read FIXATION and DNA Staining for Cell Cycle Analysis]

Category: Cell Biology and Cell Culture: Cell Cycle Protocols: G Phase Protocols

This protocol provides a method for the synchronization of a monolayer culture of CHO cells in G1 using isoleucine deprivation. Since CHO cells can also be adapted to grow in suspension culture, this procedure can be used to obtain larger quantities of cells. When isoleucine is replaced, the cells resume growth and begin to enter S phase ~4 hours later. This method arrests almost 100% of the CHO cells in G1, and upon reversal, leads to rapid recovery of cell growth and very high cell viability. - [Read G1 Synchronization of CHO Cells by Isoleucine Deprivation Protocol]

Category: Cell Biology and Cell Culture: Flow Cytometry Protocols: Cell Cycle Analysis Protocols

This protocol provides a method for the synchronization of a monolayer culture of CHO cells in G1 using isoleucine deprivation. Since CHO cells can also be adapted to grow in suspension culture, this procedure can be used to obtain larger quantities of cells. When isoleucine is replaced, the cells resume growth and begin to enter S phase ~4 hours later. This method arrests almost 100% of the CHO cells in G1, and upon reversal, leads to rapid recovery of cell growth and very high cell viability. - [Read G1 Synchronization of CHO Cells by Isoleucine Deprivation Protocol]

Category: Cell Biology and Cell Culture: Cell Cycle Protocols: G Phase Protocols

This protocol uses the plant amino acid mimosine as a G1/S synchronizing agent. Cells are first treated with excess thymidine to accumulate the majority of them at G1/S; however, some cells will have stopped growth within the S phase. Thymidine is then removed to allow all the cells to proceed completely through the S phase. Mimosine is then added to arrest the cells at the G1/S border. When mimosine is removed, cells will begin to enter S phase within about 1 hour. - [Read G1/S Phase Synchronization Using Mimosine Arrest Protocol]

Category: Cell Biology and Cell Culture: Cell Cycle Protocols: S Phase Protocols

Protocol uses the plant amino acid mimosine as a G1/S synchronizing agent. Cells are first treated with excess thymidine to accumulate the majority of them at G1/S; however, some cells will have stopped growth within the S phase. Thymidine is then removed to allow all the cells to proceed completely through the S phase. Mimosine is then added to arrest the cells at the G1/S border. When mimosine is removed, cells will begin to enter S phase within about 1 hour. - [Read G1/S Phase Synchronization Using Mimosine Arrest Protocol]

Category: Cell Biology and Cell Culture: Flow Cytometry Protocols: Cell Cycle Analysis Protocols

This protocol uses the plant amino acid mimosine as a G1/S synchronizing agent. Cells are first treated with excess thymidine to accumulate the majority of them at G1/S; however, some cells will have stopped growth within the S phase. Thymidine is then removed to allow all the cells to proceed completely through the S phase. Mimosine is then added to arrest the cells at the G1/S border. When mimosine is removed, cells will begin to enter S phase within about 1 hour. - [Read G1/S Phase Synchronization Using Mimosine Arrest Protocol]

Category: Cell Culture Protocols: Cell Immortalization Protocols

Cultivating animal cells in the laboratory is an indispensable technique for cell biologists. However, most normal primary cell lines, while faithfully reproducing the phenotype of their tissue of origin, do not grow indefinitely in culture. After a series of population doublings (the number of which varies by species, cell type, and culture conditions) primary cells enter a state where they no longer divide. - [Read Immortalization of Cells in Culture]

Category: Fungi Protocols: Aspergillus Protocols

Protocol for the transformation of Aspergillus niger. This procedure is done by first digesting the outer cell wall, forming protoplasts, and then by making holes in the membrane through which the dna can enter using calcium chloride and polyethylene glycol. Includes: Protocol for making A.niger protoplasts; Transformation; Plating. - [Read Transformation of Aspergillus niger Protocol]