An improved method for western blotting when extracting proteins from mammalian cells cultured on a collagen gel under serum-free conditions

An improved method for Southwestern blotting.

We have developed a modified Southwestern blotting technique which utilizes broad-spectrum protease inhibitors during nuclear protein extraction and a procedure for radiolabelling an oligonucleotide probe to a high specific activity. These modifications have resulted in minimal protein degradation during nuclear protein isolation and have permitted room temperature hybridizations, improving bot...

Healing Potential of Mesenchymal Stem Cells Cultured on a Collagen-Based Scaffold for Skin Regeneration

Background: Wound healing of burned skin remains a major goal in public health. Previous reports showed that the bone marrow stem cells were potent in keratinization and vascularization of full thickness skin wounds. Methods: In this study, mesenchymal stem cells were derived from rat adipose tissues and characterized by flowcytometry. Staining methods were used to evaluate their differentiatio...

Development of an Improved Sub-Molten Salt Method for Production of Potassium Titanate from Ilmenite under Industrial Conditions

Total protein extraction from cultured cells for use in electrophoresis and western blotting.

Experimentation with cultured cells often requires analyzing cellular protein extract by gel electrophoresis and immunoblotting. Traditional methods for extracting cellular proteins by homogenization or detergent solubilization usually produce protein samples that are viscous (due to the presence of DNA) and prone to degradation due to the presence of endogenous protease activity. We have devel...

Evaluation of the Criterion Stain Free Gel Imaging System for Use in Western Blotting Applications

immunodetection is performed. Use of the Criterion Stain Free system eliminates the need for the timeand reagentconsuming traditional methods such as Ponceau S staining used to asses protein transfer prior to detection in western blotting applications. We also demonstrate that, while binding of some monoclonal antibodies may be affected by the Criterion Stain Free method, we have not observed a...