What is the difference between Coomassie r250 and g250?
G-250: Which One Should You Use? Basically, both R-250 (R signifies the slightly reddish tint in the blue color of the dye) and G-250 (G signifies the greenish tint in the dye) have relatively high sensitivity and allow for easy detection (they develop intensely colored complexes upon binding with protein molecules).
How does Coomassie brilliant blue g250 work?
CBB R-250 and the dimethyl derivative CBB G-250 are disulfonated triphenylmethane dyes that stain protein bands bright blue. The dyes bind via electrostatic interaction with protonated basic amino acids (lysine, arginine, and histidine) and by hydrophobic associations with aromatic residues.
What is Coomassie brilliant blue used for?
Coomassie brilliant blue is the name of two similar triphenylmethane dyes that were developed for use in the textile industry but are now commonly used for staining proteins in analytical biochemistry.
How does Coomassie brilliant blue assay work?
This assay is performed by determining the absorbance shift of the Coomassie Brilliant Blue G-250. The amount of the complex present in the solution is used as an indicator for the protein concentration by measuring the intensity of the blue color after stabilization.
What is CBB r250?
Coomassie R-250 and G-250 dyes are two chemical forms of a disulfonated triphenylmethane compound that is commonly used as the basis of stains for detection of proteins in gel electrophoresis and Bradford-type assay reagents for protein quantitation.
What is the purpose of Coomassie staining?
Coomassie Blue stain is used to stain the protein bands in polyacrylamide gels. One common way to use it is to dissolve the dye in a mixture of methanol, acetic acid, and water. This stain will permeate the gel, stain the protein, and also fix the protein in place.
What does a Coomassie stain tell you?
In acidic conditions, Coomassie dye binds to basic and hydrophobic residues of proteins, changing in color from a dull reddish-brown to intense blue. As with all staining methods, Coomassie staining detects some proteins better than others, based on the chemistry of action and differences in protein composition.
How does Coomassie Blue stain bind to proteins?
In the staining reaction, the Coomassie dye binds to proteins through ionic interactions between sulfonic acid groups and positive protein amine groups through Van der Waals attractions.
How does the Coomassie Brilliant Blue G-250 dye binds to a protein?
Does Coomassie Blue stain all proteins?
The most common used protein stain is Coomassie Blue staining, which is based on the binding of Coomassie Brilliant Blue, which binds non-specifically to virtually all proteins.
How does Coomassie stain work and why is it useful?
Which amino acid does Coomassie bind to?
In acidic conditions, the dye binds to proteins primarily through basic amino acids (primarily arginine, lysine and histidine), and the number of coomassie dye ligands bound to each protein molecule is approximately proportional to the number of positive charges found on the protein.
What is Coomassie Brilliant Blue G-250?
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 do you use Coomassie Brilliant Blue R-250 stain?
Use this Coomassie brilliant blue R-250 solution to stain proteins in SDS-PAGE gels. Step 1: Stain gels for 1–2 hours with gentle agitation. Stain overnight or longer if needed. Step 2: Destain gels for 2 hours. Change destaining solution ( 1610438) multiple times (e.g., 4 washes x 30 min) until the background is less dark.
What is the difference between R250 and G250 Coomassie dye?
The R-250 (red-tinted) form lacks two methyl groups that are present in the G-250 (green-tinted) form, which is also called colloidal coomassie dye. Typically, coomassie gel stains and protein assay reagents are formulated as very acidic solutions in 25 to 50% methanol.
How does Coomassie dye bind to proteins?
In acidic conditions, the dye binds to proteins primarily through basic amino acids (primarily arginine, lysine and histidine), and the number of coomassie dye ligands bound to each protein molecule is approximately proportional to the number of positive charges found on the protein.