Preparation of staining solution CBB R mg was dissolved in ml of distilled water by stirring for 2—4 h. Gel electrophoresis The stacking and separating gels used were 3. Results and Discussion Sensitivity The Neuhoff's Chinacolloidal Coomassie Blue G staining has a detection limit of approximately 10 ng of protein per spot [16].
Open in a separate window. Figure 1. Sensitivity of the described procedure. Different acids To evaluate whether the acids have any effects on the protocol, we used different acids, including hydrochloric acid, phosphoric acid, and acetic acid, to prepare the staining solution in the experiment.
Figure 2. Gels stained with different staining solutions. Different time and temperature Temperature is a very important factor for protein detection in the protocol. Figure 3. Gels stained at different time periods. Water purity Washing before staining is necessary to prepare a clean background. Figure 4. Gel washing with different types of water. Footnotes Competing Interests: The authors have declared that no competing interests exist.
References 1. Walker J, editor. Humana Press Totowa. Elevated expression of membrane-type 1 and 3 matrix metalloproteinases in rat vascular smooth muscle cells activated by arterial injury. Lab Invest. Isolation, purification, characterization and glycan-binding profile of a d-galactoside specific lectin from the marine sponge, Halichondria okadai.
Glycan-binding profile of a D-galactose binding lectin purified from the annelid, Perinereis nuntia ver. An improved formulation of SYPRO Ruby protein gel stain: Comparison with the original formulation and with a ruthenium II tris bathophenanthroline disulfonate formulation. Blue silver: A very sensitive colloidal Coomassie G staining for proteome analysis. Two new staining procedures for quantitative estimation of proteins on electrophoretic strips.
Biochim Biophys Acta. Ultrasensitive stain for proteins in polyacrylamide gels shows reqional variation in cerebrospinal fluid proteins. Weber K, Osborn M. The reliability of molecular weight determinations by dodecyl sulfate-polyacrylamide gel electrophoresis.
Bid Chem. Bennet J, Scott J. Quantitative staining of fraction 1 protein in polyacrylamide gels using Coomassie Brilliant Blue. Anal Biochem. Staining and destaining polyacrylamide gels: a comparison of Coomassie Blue and fast green protein dyes. Anal Sci. CBB staining protocol with higher sensitivity and mass spectrometric compatibility.
A modified Coomassie blue staining of proteins in polyacrylamide gels with Bismark brown R. Anal Bioanal Chem. Dyballa N, Metzger S. Fast and sensitive colloidal coomassie G staining for proteins in polyacrylamide gels. J Vis Exp. Fast protein staining in sodium dodecyl sulfate polyacrylamide gel using counter ion-dyes, Coomassie brilliant blue R and neutral red. Arch Pharm Res. Proteins with unusually high proportions of ring-y amino acids tend to stain better. An example is BSA bovine serum albumin , which recruits twice as many treasure hunters per weight of protein than your average protein.
Speaking of weight, different proteins have different weights because they have different s of amino acid letters. That number might seem random, but it's Avogadro's number and it's defined as the number of atoms in 12 grams of carbon the main isotope version of carbon. Because molecules are really tiny. These terms are often used interchangeably, but, technically Let's start with MW units: Dalton, unified atomic mass unit u , and atomic mass unit amu are, practically speaking, the same.
We usually use Da in biochemistry to talk about the sizes of proteins and stuff. But to make things more confusing Then they redefined things a little so they're a tiny bit different, but that only really matters for physic-y stuff and for the stuff we're talking about, we can just consider them the same and you'll often see all these terms interchanged! When I purify proteins, I'm glad to get 5mg at the end of the process!
So your bands would look darker for bigger proteins than smaller proteins if you ran the same of protein molecules of each. CBB can also bind SDS that detergent we used earlier to solubilize and negatively-charge the proteins which can mess up results. So you want to wash your gel in water before staining to remove the SDS.
The classical CBB method goes something like this apologies for scribbled-note-like formatting …. So how do you get enough to stain the protein? The result? Bio-Rad Products Back. Life Science Education Explore all. About the Program Explore all. Corporate Explore all. About Bio-Rad Back. About Bio-Rad Explore all.
Investor Relations Explore all. Coomassie Stains. Bio-Safe Coomassie Gel. Bio-Rad offers Coomassie stains in three major formats. Description Ordering Documents. Coomassie G is more sensitive Coomassie R provides better resolution and lower cost. Introduction to Polyacrylamide Gels. Imaging and Analysis of 2-D Electrophoresis Gels.
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