decreased Protocols

Category: Cell Biology and Cell Culture: Apoptosis Protocols: Apoptosis Analysis

Analysis of DNA Fragmentation Using the JAM Assay. By Shailaja Kasibhatla et al., The JAM assay is based on labeling nuclear DNA of cycling cells with [3H]thymidine and harvesting samples on glass fiber filters. Apoptosis will generate DNA fragments small enough to pass through the glass fiber filter, resulting in decreased radioactivity of the particular sample. Cell-mediated cytotoxicity or cell killing mediated by cytotoxic T lymphocytes (CTL) can also be measured by this technique. - [Read Analysis Of DNA Fragmentation Using The JAM Assay (Subscription Required)]

Category: Mouse Protocols

Mice fed with the cytohesin inhibitor SecinH3 for two days develop hepatic insulin resistance that can be identified by reduced liver glycogen levels, increased serum insulin and ketone body levels and decreased serum non-esterified fatty acid. To confirm the presence and identity of SecinH3 in mouse liver, we extracted the compound from liver homogenates with chloroform and identified it by LC/MS. - [Read Extraction of the SecinH3 from Mouse Liver Protocol]

Category: Staining Protocols: Gram Staining Protocols

Gram-positive and Gram-negative organisms form a complex of crystal violet and iodine within the bacterial cell during the Gram-staining procedure. Gm+ organisms are thought to resist decolorization by alcohol or acetone because cell wall permeability is markedly decreased when it is dehydrated by these solvents. Thus, the dye complex is entrapped within the cell, resist being washed out by the solvents, and Gm+ bacteria remain purple following this differential stain. - [Read Gram Staining Protocol]

Category: Pharmacology and Toxicology Protocols: Cytotoxicity Protocols: Cell Specific Cytotoxicity Assays Protocols

The basis of this procedure is that two specific cell type preparations may be isolated, exposed separately to various compounds over a range of concentrations, and the cytotoxicity of these determined.  Parameters deemed indicative of a cytotoxic effect include a reduction in de novo protein synthesis and decreased glucose and fatty acid metabolism.  A cytotoxic effect may indicate that a chemical is likely to be nephrotoxic in vivo. - [Read Isolated Rat Glomeruli and Proximal Tubules]

Category: DNA Protocols: cDNA Library Protocols: cDNA Library Construction Protocols

PCR can be used to identify rare DNA sequences in DNA libraries by increasing the abundance of a particular sequence. This is accomplished by subdividing the original library into pools of decreased complexity and screening each pool or group of pools for a given DNA sequence. - [Read PCR-Based Screening of DNA Libraries Protocol]

Category: PCR Protocols

PCR can be used to identify rare DNA sequences in DNA libraries by increasing the abundance of a particular sequence.  This is accomplished by subdividing the original library into pools of decreased complexity and screening each pool or group of pools for a given DNA sequence.  A pool that contains the desired clone is subsequently subdivided into smaller pools, each of which is screened using the same PCR protocol that was used for the primary screen. - [Read PCR-Based Screening of DNA Libraries Protocol]