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Polysorbate 20 is also known as Tween 20, a commercial brand name. It is a common detergent used in many buffers for washing nitrocellulose membrane in western blotting and microtiter plate wells in ELISA assays. Tris is a buffer that maintains a pH of 7–9.2.
The western blot method is composed of gel electrophoresis to separate native proteins by 3-D structure or denatured proteins by the length of the polypeptide, followed by an electrophoretic transfer onto a membrane (mostly PVDF or nitrocellulose) and an immunostaining procedure to visualize a certain protein on the blot membrane.
Following electrophoresis, a standard tank or semi-dry blotting transfer system is set up. A stack is put together in the following order from cathode to anode: sponge | three sheets of filter paper soaked in transfer buffer | gel | PVDF or nitrocellulose membrane | three sheets of filter paper soaked in transfer buffer | sponge.
These can be transferred onto a nitrocellulose or PVDF membrane to be probed with antibodies and corresponding markers, such as in a western blot. Typically resolving gels are made in 6%, 8%, 10%, 12% or 15%. Stacking gel (5%) is poured on top of the resolving gel and a gel comb (which forms the wells and defines the lanes where proteins ...
Amido black 10B is an amino acid staining azo dye used in biochemical research to stain for total protein on transferred membrane blots, such as the western blot. [2] It is also used in criminal investigations to detect blood present with latent fingerprints - it stains the proteins in blood a blue-black color. [3]
Ponceau S, Acid Red 112, or C.I. 27195 (systematic name: 3-hydroxy-4-(2-sulfo-4-[4-sulfophenylazo]phenylazo)-2,7-naphthalenedisulfonic acid sodium salt [1]) is a sodium salt of a diazo dye of a light red color, that may be used to prepare a stain for rapid reversible detection of protein bands on nitrocellulose or polyvinylidene fluoride (PVDF) membranes (western blotting), as well as on ...
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