Polyvinylidene fluoride Polyvinylidene Fluoride, or PVDF is a highly non-reactive and pureIt is also known as KYNAR, HYLAR or SYGEF.

A fluoropolymer is a fluorocarbon based polymer with multiple strong carbonfluorine bonds. It is characterized by a high resistance to solvents, acids, and bases. *A thermoplastic is a polymer that turns to a liquid when heated and freezes to a very glassy state when cooled sufficiently. PVDF is a specialty plastic material in the fluoropolymer family; it is used generally in applications requiring the highest purity, strength, and resistance to solvents, acids, bases and heat and low smoke generation during a fire event. Compared to other fluoropolymers, it has an easier melt process because of its relatively low melting point of around 177C. It has a low density (1.78) and low cost compared to the other fluoropolymers. It is available as piping products, sheet, tubing, films, plate and an insulator for premium wire. It can be injected, molded or welded and is commonly used in the chemical, semiconductor, medical and defense industries, as well as in lithium ion batteries. A fine powder grade, KYNAR 500 PVDF or HYLAR 5000 PVDF, is also used as the principal ingredient of high-end paints for metals. These PVDF paints have extremely good gloss and color retention, and they are in use on many prominent buildings around the world, e.g. the Petronas Towers in Malaysia and Taipei 101 in Taiwan, as well as on commercial and residential metal roofing. PPG Industries, Inc. is a well-known supplier of such PVDF-containing coatings. PVDF membranes are used for western blots for immobilization of proteins, due to its non-specific affinity for amino acids.

Analytical technique
From Wikipedia, the free encyclopedia

Jump to: navigation, search An analytical technique is a method that is used to determine the concentration of a chemical compound or chemical element. There are a wide variety of techniques used for analysis, from simple weighing (gravimetric) to titrations (titrimetric)to very advanced techniques using highly specialized instrumentation. The most common techniques used in analytical chemistry are the following:

titrimetry, based on the quantity of reagent needed to react with the analyte Electroanalytical techniques, including potentiometry and voltammetry Spectroscopy, based on the interaction of the analyte with electromagnetic radiation Chromatography, in which the analyte is separated from the rest of the sample so that it may be measured without interference from other compounds

There are many more techniques that have specialized applications, and within each major analytical technique there are many applications and variations of the general techniques.
Gel electrophoresis is a technique used for the separation of deoxyribonucleic acid (DNA), ribonucleic acid (RNA), or protein molecules using an electric current applied to a gel matrix.[1] It is usually performed for analytical purposes, but may be used as a preparative technique prior to use of other methods such as mass spectrometry, RFLP, PCR, cloning, DNA sequencing, or Southern blotting for further characterization. Nitrocellulose (also: cellulose nitrate, flash paper) is a highly flammable compound formed by nitrating cellulose through exposure to nitric acid or

another powerful nitrating agent. When used as a propellant or low-order explosive, it is also known as guncotton.

A nitrocellulose slide, nitrocellulose membrane or nitrocellulose paper is a sticky membrane used for immobilizing nucleic acids in Southern blots and northern blots. It is also used for immobilization of proteins in western blots, due to its nonspecific affinity for amino acids. Nitrocellulose is widely used as support in diagnostic tests where antigen-antibody binding occur, e.g., pregnancy tests, U-Albumin tests and CRP. Glycine and chloride ions make protein transfer more efficient. When dissolved in ether or other organic solvents, the solution is called collodion, which has been used as a wound dressing and carrier of topical medications since the U.S. Civil War. To this day, it is used in Compound W Wart Remover as a carrier of salicylic acid, the active ingredient. Collodion was also used as the carrier for silver salts in some very early photographic emulsions, particularly spread in thin layers on glass plates. Magician's flash paper, sheets of paper or cloth made from nitrocellulose, which burn almost instantly, with a bright flash, and leave no ash. Radon tests for alpha track etches Nitrocellulose lacquer was used as a finish on guitars for most of the 20th century and is still used on some current applications. Manufactured by (among others) Dupont, the paint was also used on automobiles sharing the same color codes as many guitars including Fender and Gibson brands.[5] Nitrocellulose lacquer is also used as an aircraft dope, painted onto fabric-covered aircraft to tauten and provide protection to the material. As a transportation medium for one-time pads, thus making the disposal of the pad complete, secure, and efficient. Depending on the manufacturing process, nitrocellulose is esterified to varying degrees. Table tennis balls, guitar picks and some photographic films have a fairly low esterification level

and burn comparatively slowly with some charred residue. See celluloid. Due to it's explosive nature, not all applications of nitrocellulose were successful. In 1869, with elephants having been poached to near extinction, the billiards industry offered a $10,000 prize to whomever came up with the best replacement for ivory billiard balls. John Wesley Hyatt created the winning replacement which he coated with a new material he discovered called camphored nitrocellulosethe first thermoplastic, better known as celluloid. The invention enjoyed a brief popularity, but the Hyatt balls were extremely flammable, and sometimes portions of the outer shell would explode upon impact. An owner of a billiard saloon in Colorado wrote to Hyatt about the explosive tendencies, saying that he did not mind very much personally but for the fact that every man in his saloon immediately pulled a gun at the sound.[6][7] The process used by Hyatt to manufacture the billiard balls, (US Patent 239,792, 1881) involved placing the mass of nitrocellulose in a rubber bag, which was then placed in a cylinder of liquid and heated. Pressure was applied to the liquid in the cylinder, which resulted in a uniform compression on the nitrocellulose mass, compressing it into an uniform sphere as the heat vaporizes the solvents. The ball then cooled, and turned to make a uniform sphere. Ironically, in light of the explosive results, this process was called the "Hyatt Gun Method".[8]

Western blot
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The western blot (alternatively, immunoblot) is an analytical technique used to detect specific proteins in a given sample of tissue homogenate or extract. It uses gel electrophoresis to separate native or denatured proteins by the length of the polypeptide (denaturing conditions) or by the 3-D structure of the protein (native/ non-

denaturing conditions). The proteins are then transferred to a membrane (typically nitrocellulose or PVDF), where they are probed (detected) using antibodies specific to the target protein.[1] [2] There are now many reagent companies that specialize in providing antibodies (both monoclonal and polyclonal antibodies) against tens of thousands of different proteins[3]. Commercial antibodies can be expensive, although the unbound antibody can be reused between experiments. This method is used in the fields of molecular biology, biochemistry, immunogenetics and other molecular biology disciplines. Other related techniques include using antibodies to detect proteins in tissues and cells by immunostaining and enzyme-linked immunosorbent assay (ELISA). The method originated from the laboratory of George Stark at Stanford. The name western blot was given to the technique by W. Neal Burnette[4] and is a play on the name Southern blot, a technique for DNA detection developed earlier by Edwin Southern. Detection of RNA is termed northern blotting and the detection of posttranslational modification of protein is termed Eastern blotting.

Contents [hide]

1 Steps in a western blot

o o o o o

1.1 Tissue preparation 1.2 Gel electrophoresis 1.3 Transfer 1.4 Blocking 1.5 Detection

1.5.1 Two step 1.5.2 One step 1.6.1 Colorimetric detection 1.6.2 Chemiluminescent detection 1.6.3 Radioactive detection 1.6.4 Fluorescent detection

1.6 Analysis

1.7 Secondary probing

2 2-D Gel Electrophoresis 3 Medical diagnostic applications 4 Protocols 5 References 6 See also
o

6.1 Related links

[edit] Steps in a western blot

[edit] Tissue preparation

Samples may be taken from whole tissue or from cell culture. In most cases, solid tissues are first broken down mechanically using a blender (for larger sample volumes), using a homogenizer (smaller volumes), or by sonication. Cells may also be broken open by one of the above mechanical methods. However, it should be noted that bacteria, virus or environmental samples can be the source of protein and thus Western blotting is not restricted to cellular studies only.

Assorted detergents, salts, and buffers may be employed to encourage lysis of cells and to solubilize proteins. Protease and phosphatase inhibitors are often added to prevent the digestion of the sample by its own enzymes. Tissue preparation is often done at cold temperatures to avoid protein denaturing. A combination of biochemical and mechanical techniques including various types of filtration and centrifugation can be used to separate different cell compartments and organelles.

Gel electrophoresis
Main article: Gel electrophoresis

Immunoblot (Western blot) analysis of proteins separated by SDS-PAGE gradientgel electrophoresis.[5]

The proteins of the sample are s