Two-Dimensional Gel Electrophoresis: Vertical Isoelectric Focusing.

Two-dimensional gel electrophoresis is one of the most powerful tools for separating proteins based on their size and charge. Two-dimensional gel electrophoresis (2-DE) is very useful to separate two proteins with identical molecular weights but different charges, which cannot be achieved with just sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE). Here, a simpler and easier version of 2-DE is presented which is also faster than all the currently available techniques.

Paper Adsorbents Remove Coomassie Blue from Gel Destain and Used Gel Stain in an Environment-Friendly Manner.

Coomassie Brilliant Blue (CBB), used to stain protein gels, is known to be toxic. Therefore, laboratories do not discard used CBB into the sink owing to the possibility of it contaminating drinking water supplies. We tested the ability of various paper adsorbents to adsorb CBB released from gels during destaining.

Curcumin/Turmeric as an Environment-Friendly Protein Gel Stain.

Curcumin, the main curcuminoid in food spice turmeric, is insoluble in water at room temperature. We showed that curcumin can be solubilized in water with the application of heat (100 °C). Here we demonstrate that heat-solubilized curcumin can serve as a nontoxic and environment-friendly fluorescent/colorimetric reversible protein stain.

Immunoblotting of Antigens: Whole, Strip, and New-Line Nitrocellulose Membrane Immunoblotting Using the Chemiluminescence Technique.

Antigen detection is a well-known tool in the scientific world that is used by clinicians and researchers to detect specific antigens in diagnosing diseases or for other medical/environmental discoveries. Antigen detection is introduced in various forms over the past decades. These techniques are often evaluated by their sensitivity, accuracy, and ease of use.

A miniaturized blotting system for simultaneous detection of different autoantibodies.

Sera of tumor patients frequently contain autoantibodies to tumor associated antigens. Here we describe a miniaturized immunoblot platform allowing us to screen sera of patients for the presence of autoantibodies to ten autoantigens in parallel.

Induction of anti-Ro60/anti-La by immunisation with spectrin and induction of anti-spectrin by immunisation with Ro60 and 4-hydroxy-2-nonenal-modified Ro60 immunisation.

Objectives: The Ro ribonucleoprotein particle, targeted in systemic lupus erythematosus (SLE) and Sjögren's syndrome (SS), includes Ro60 (SSA) and La (SSA) autoantigens. Anti-Ro60 occurs in SLE and SS. The importance of α-fodrin and spectrin as well as anti-Ro and anti-fodrin/spectrin antibodies in SS and SLE, led us to hypothesise that rabbit immunisation with Ro60 or 4-hydroxy-2-nonenal-modified Ro60 would induce anti-spectrin.

Coomassie Brilliant Blue removal/disposal from gel destain and used gel stain in an environment-friendly manner.

Toxic reagents are employed to destain Coomassie Brilliant Blue (CBB) stained gels. We tested the efficacy of various paper adsorbents in adsorbing CBB released from gels during destaining. Kimwipes were the most efficient, followed by Teri towels, multifold towels, and Whatman (numbers 1 and 3) filter papers.

Spicy SDS-PAGE gels: curcumin/turmeric as an environment-friendly protein stain.

Gel proteins are commonly stained with calorimetric/fluorescent dyes. Here, we demonstrate that heat-solubilized curcumin can serve as a nontoxic and environment-friendly fluorescent/colorimetric reversible protein stain. Curcumin, the yellow pigment found in the rhizomes of the perennial herb Curcuma longa (turmeric), is insoluble in aqueous solvents.

Two-dimensional gel electrophoresis: vertical isoelectric focusing.

Two-dimensional gel electrophoresis (2-DE) is one of the most powerful tools for separating proteins based on their size and charge. 2-DE is very useful to separate two proteins with identical molecular weights but different charges, which cannot be achieved with just sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE). Here, a simpler and easier version of 2-DE is presented which is also faster than all the currently available techniques.

An overview of Western blotting for determining antibody specificities for immunohistochemistry.

Despite its overall simplicity, protein blotting or Western blotting has been proven to be a powerful procedure for the immunodetection of proteins, especially those that are of low abundance, following electrophoresis. The usefulness of this procedure stems from its ability to provide simultaneous resolution of multiple immunogenic antigens within a sample for detection by specific antibodies. Protein blotting has evolved greatly since its inception and researchers have a variety of ways and means to carry out this transfer.

Degree of modification of Ro60 by the lipid peroxidation by-product 4-hydroxy-2-nonenal may differentially induce Sjögren syndrome or systemic lupus erythematosus in BALB/c mice.

Our previous work showed that immunization of rabbits with 4-hydroxy-2-nonenal-modified Ro60 (HNE-Ro60) accelerates autoimmunity. We extended this model into mice, hypothesizing that the severity of autoimmunity would be dependent on the degree of HNE modification of Ro60. Five groups of BALB/c mice (10/group) were used.

Environmentally safe removal/disposal of Coomassie Brilliant Blue from gel destain and used gel stain.

Gel destaining following Coomassie Brilliant Blue (CBB) staining involves the use of toxic reagents. Here we demonstrate the efficacy of various paper adsorbents in adsorbing CBB. Kimwipes adsorbed the best, followed by Teri towels, multifold towels, and Whatman numbers 1 and 3 filter papers.

Problems with multiple use of transfer buffer in protein electrophoretic transfer.

Two-dimensional gel electrophoresis (2DE) and SDS-PAGE are the two most useful methods in protein separation. Proteins separated by 2DE or SDS-PAGE are usually transferred to membranes using a variety of methods, such as electrophoretic transfer, heat-mediated transfer, or nonelectrophoretic transfer, for specific protein detection and/or analysis. In a recent study, Pettegrew et al.

Whole and strip nitrocellulose membrane as well as a new-line-immunoblotting of antigen using the chemiluminescence technique.

There are a number of techniques in the scientific world that researchers use to detect specific antigens. One such technique that has provided many advantages over typical immunochemical staining is chemiluminescence. The emission of visible radiation by compounds once exposed to sunlight has been known for centuries and currently is the main principle for chemiluminescence.

A miniaturized blotting system for simultaneous detecting of different autoantibodies.

Sera of tumor patients frequently contain autoantibodies to tumor-associated antigens. Here we describe a miniaturized immunoblot platform allowing us to screen sera of patients for the presence of autoantibodies to ten autoantigens in parallel.

Awaken olfactory receptors of humans and experimental animals by coffee odourants to induce appetite.

Smell and its mechanism has been of interest to scientists for many years. Smell, not only provides a sensual pleasure of food and perfumes for humans but also reminds us of past memories, thoughts, locations and finally warns of dangers such as fire. One of the uses of coffee beans is on perfume counters, enabling people to distinguish between perfume fragrances.

Four men in a space station - To say nothing of the cow! The quest for finding respite and work in the ultimate frontier.

Fed up with life on earth, four scientists attempt to make it to space to live in the International Space Station (ISS) and carry out experiments. The difficulties in getting selected by NASA, the rigourous training to fly and the risks of the journey to life and health are the rate limiting steps in their quest. They propose commercialization of space and also ferrying cows to space for food as well as generation of biogas.