Polyacrylamide gels are typically formed by polymerization of the monomer acrylamide crosslinked to the co-monomer, N,N’-methylenebis-acrylamide, commonly called BIS. This process is a freeradical polymerization that requires an initiator, usually ammonium persulfate, and a catalyst like N,N,N’,N’-tetraethylmethylenediamine (TEMED). A distinct advantage of polyacrylamide gel systems is that by ...

Electrophoresis is widely used as a separative technique, particularly in protein chemistry. In some respects gelatinous media have an advantage over others as support media for electrophoresis, perhaps most obviously when a gel matrix of molecular dimensions is employed, allowing the simultaneous use of two properties for separation of the particles of interest-size and surface charge density....

OBJECTIVE To identify the relative abundance of proteins in pooled reactive oxygen species (ROS)-positive (ROS+) and ROS-negative (ROS-) semen samples with the use of two-dimensional differential in-gel electrophoresis (2D-DIGE). DESIGN Spermatozoa suspensions from ROS+ and ROS- groups by 2D-DIGE analysis. SETTING Tertiary hospital. PATIENT(S) 20 donors and 32 infertile men. INTERVENTIO...

Coxiella burnetii is an obligate intracellular bacterial pathogen and the causative agent of Q fever. Chronic Q fever can produce debilitating fatigue and C. burnetii is considered a significant bioterror threat. C. burnetii occupies the monocyte phagolysosome and although prior work has explained features of the host-pathogen interaction, many aspects are still poorly understood. We have condu...

The Streptomyces griseus 70S ribosome fraction was analyzed by radical-free and highly reducing two-dimensional (RFHR 2D) gel electrophoresis and mass spectrometry. Among the 60 putative ribosomal proteins that are encoded by the S. griseus genome, 48 were identified in the 70S ribosome fraction prepared from mycelia grown in liquid culture for 12, 36, and 48 h. Ribosomal protein S3 was detecte...

INTRODUCTION Alkaline agarose gels are run at a pH that is sufficiently high to denature double-stranded DNA. The denatured DNA is maintained in a single-stranded state and migrates through the alkaline gel as a function of its size. Alkaline agarose gels are used chiefly to measure the size of first and second strands of cDNA (Construction of cDNA Libraries Stage 1: Synthesis of First-strand c...

U.V. scanning may readily be performed on unstained agar-gel strips in the dry state. Polyacrylamide gels tend to become brittle if dried; whether they can be reliably scanned in the undried state, as unstained strips or columns, depends on whether there are troubles due to deformation of the pattern when the gel is manipulated, or to optical opacity. With proteins, if not with RNA, u.v.-sensit...