What is Coomassie Brilliant Blue R-250?
Coomassie Brilliant Blue R-250 (CBBR) is a member of the Coomassie family and is used extensively as an analytical dye in SDS-PAGE. CBBR can also be used in the Bradford Assay to help quantify the protein concentration in a mixture, like cell lysates.
What is Coomassie Blue How does it function?
Description. Coomassie blue dyes are a family of dyes commonly used to stain proteins in SDS-PAGE gels. The gels are soaked in dye, and excess stain is then eluted with a solvent (“destaining”). This treatment allows the visualization of proteins as blue bands on a clear background.
How do you cook Coomassie Brilliant Blue G 250?
Dissolve 0.25 g of Coomassie Brilliant Blue R-250 in 90 ml of methanol:H2O (1:1, v/v) and 10 ml of glacial acetic acid. Filter the solution through a Whatman No. 1 filter to remove any particulate matter.
What is Coomassie brilliant blue used for?
Coomassie brilliant blue (CBB) stain is a widely used method for routine visualization of proteins separated on polyacrylamide gels. It is an organic dye that makes complexes with basic amino acids, such as lysine, histidine, tyrosine, and arginine.
What does Coomassie Brilliant Blue bind to?
CBB binds to basic amino acids by a combination of hydrophobic interactions and heteropolar bonding, forcing proteins to move toward the anode, independently of their intrinsic pI.
What wavelength does Coomassie blue maximally absorb at?
465 nm
There is a shift in the peak absorbance of the dye when it binds to a protein. Unbound Coomassie Blue absorbs light maximally at a wavelength of 465 nm, while the absorption maximum is at 595 nm when the dye is bound to protein.
What amino acids does Coomassie G-250 bind to?
In acidic conditions, Coomassie dye primarily binds basic amino acids (arginine, lysine and histidine).
How does Coomassie Brilliant Blue G-250 bind to proteins?
When Coomassie Brilliant Blue G-250 binds to proteins in acid solution, it has an absorbance shift from 465 nm to 595 nm. The absorbance data can then be used in Beer’s law to determine protein concentration and ultimately the actual amount of protein in a given solution.
Where can I buy Coomassie Brilliant Blue G-250 dye?
ERR#1200 – This item cannot be ordered online. Order requests are processed and shipped by your local distributor. Learn more Thermo Scientific Pierce Coomassie Brilliant Blue G-250 is one of the most common forms of coomassie dye, which is a key component of various colorimetric protein gel stains.
How is Coomassie Brilliant Blue used to stain proteins?
In classical protein staining protocols using Coomassie Brilliant Blue (CBB), solutions with high contents of toxic and flammable organic solvents (Methanol, Ethanol or 2-Propanol) and acetic acid are used for fixation, staining and destaining of proteins in a gel after SDS-PAGE.
Which is more sensitive, Coomassie blue or G250?
Coomassie Blue is widely used in visualizing proteins separated by either agarose or acrylamide gel electrophoresis. The more popular is Coomassie R250 (Reddish tinted blue) for electrophoresis (more sensitive: can detect as little as 0.1 µg of protein), and Coomassie G250 (Greenish tinted blue) for protein assay in solutions (because it is more
