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Topic: Coomassie blue

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	Olympus MIC-D: Polarized Light Gallery - Brilliant Blue (Coomassie Blue)
		Coomassie blue binds to proteins in an approximately stoichometric manner, meaning that when the relative amounts of protein need to be assessed, densitometry of the stained protein will provide data.
		Other uses for brilliant blue dyes are detection of lipids in thin layer chromatography, for detection of proteins in solution and on cellulose acetate plates, and as a traceable dye for following water uptake in the roots of plants.
		To a chemist, brilliant blue is actually a diammonium salt known also as acid blue or a disodium salt, sometimes called alphazurine, featuring 37 carbon atoms, 34 hydrogens, 2 nitrogens, 2 sodiums, 9 oxygens, and 3 sulfurs per molecule.
	www.olympusmicro.com /micd/galleries/polarized/brilliantblue.html (459 words)

	Bradford Assay, Development, Tobacco Hornworm, Research Link 2000
		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.
		Coomassie blue (the dye in the Bradford Reagent), phosphoric acid, and methanol are very harmful when swallowed, absorbed through the skin or splashed in the eyes.
		Coomassie blue, phosphoric acid, and methanol are components of the Bradford Reagent.
	www.acad.carleton.edu /curricular/BIOL/resources/rlink/lab2p2.html (964 words)

	Coomassie - Wikipedia, the free encyclopedia
		Coomassie (also known as Brilliant Blue, Brilliant Blue G, Acid Blue 90, C.I., or Brilliant Blue G 250) is a blue dye commonly used in sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE).
		Coomassie dye is an integral component of the Bradford Method for determining protein concentration in a solution.
		Like many dyes, Coomassie blue dye is named after an African locale, in this case the city of Kumasi, now in Ghana.
	en.wikipedia.org /wiki/Coomassie (321 words)

	The Scientist : Dye Hard: Protein Gel Staining Products (Site not responding. Last check: 2007-10-11)
		Coomassie Blue was indeed originally manufactured as an acid wool dye, or a dye that stains wool under acidic conditions (C.M. Wilson, Methods in Enzymology, 91:236-47, 1983).
		Known as Coomassie Blue G-250 and Coomassie Blue R-250, the dyes stain with green and reddish tints, respectively.
		The Colloidal Coomassie G-250 Stain from Zaxis is as sensitive as the Merril Silver Stain procedure (described later) and is highly quantitative with minimal background.
	www.the-scientist.com /article/display/18035 (4631 words)

	PAGE GEL STAINING (Site not responding. Last check: 2007-10-11)
		For optimal visualization of proteins on Coomassie blue stained gels, additional protein needs to be loaded relative to that used for Western blots.
		Coomassie Blue staining is reported to be 3 times more sensitive than Fast Green and 6 more sensitive than Amido Black.
		Coomassie blue is used to stain the gel, not a Western blot membrane.
	omrf.ouhsc.edu /~frank/SILVER.html (867 words)

	Experiment 8
		The binding of the Coomassie Brilliant Blue G-250 dye to proteins causes a shift in the dye's maximum absorbance wavelength.
		Coomassie is a hydrophobic dye that changes to an even deeper blue when it binds to a protein.
		Coomassie staining will detect a single protein band that is present at between 0.2 and 20 µg.
	www-class.unl.edu /bioc433/experiment8.htm (3976 words)

	FAQ
		Coomassie blue-stained 2D gel (~300 ug, left) and PVDF blot (~100 ug protein, right) of Eukaryotic cell line Tera I. Conditions: pH 3.5-10 ampholines, 8% acrylamide slab gels.
		Protein concentrations for complex samples such as cell lysates should be about 5 mg/ml for Coomassie blue and 1 mg/ml for silver staining since the maximum volume that can be loaded is 50 ul.
		The rule of thumb is that a polypeptide spot must be fairly dark with Coomassie blue-staining on a 2D gel to be in range for sequencing.
	www.kendricklabs.com /example.htm (1912 words)

	W.M. Keck Facility at Yale - Sample Preparation for In Situ Enzymatic Digestion in SDS Page Gels
		Since Coomassie Blue staining intensity varies by at least two-fold (depending upon the protein) it is important that more than one standard protein be run on the same gel as the sample and that an average staining intensity be used to estimate the amount of protein in the sample.
		Since this estimate may well determine the procedures that may be taken to either identify the protein or to obtain internal Edman sequences, it is extremely important that the estimate of the amount of protein submitted be as accurate as reasonably possible.
		However, regardless of the [Coomassie Blue], the gel should be exposed to 10% acetic acid, 50% methanol for a total (stain plus destain) period of at least 3 hours (with shaking and at least three solvent changes) to ensure adequate removal of SDS.
	keck.med.yale.edu /prochem/stain.htm (1098 words)

	HistoSoft » Bradford Assay » Bradford Assay Background (Site not responding. Last check: 2007-10-11)
		Free, or unbound, Coomassie molecules are most readily detected around 470 nm while bound molecules are most readily detected at 595 nm.
		As the Coomassie preferentially binds to select amino acids and changes from a cationic (+) state to an anionic (-) one, its bound condition is best detected at the maximal absorbance spectrum at 595 nm.
		The elevated absorbance at this wavelength reflects the increased binding of protein-Coomassie and that relationship is proportional - the higher the value at this optimal wavelength, the higher the amino acid (or protein) content in a sample.
	www.histosoft.com /services/background/ba/baback.html (1206 words)

	WesternBlotting.org - Coomassie Brilliant Blue Protein Stain
		Coomassie Brilliant Blue R250, binds nonspecifically to virtually all proteins.
		Coomassie Blue staining is less sensitive than silver staining, but effective nonetheless - in addition to being easy to perform.
		Excess Coomassie stain will be washed out during destaining (30 min).
	www.westernblotting.org /Coomassie.htm (118 words)

	Price Guide
		One Coomassie blue stained, dried 2-D gel of dried dimensions 16 cm wide x 13 cm long, is returned for each sample along with a pH gradient plot and a method description sheet suitable for reports or publications.
		The Coomassie pattern shows that most of the protein in the sample is from serum proteins, suggesting that the cell pellet was prepared without washing and substantially diluted.
		None of the proteins identified by the Western blot are visible on the Coomassie blue gel; for this sample the proteins would have to be immunoprecipitated to obtain enough material for mass spectrometry.
	www.kendricklabs.com /pric.htm (4147 words)

	NIBSC CJD Resource Centre-NPOR00121 Coomassie gel
		Samples were run on a gel and coomassie brilliant blue stained to demonstrate protein purity.
		Samples were prepared as serial dilutions in PBS (phosphate buffered saline).An equal volume of 2X denaturation gel loading buffer was added and samples were heated to 95°C for 5 mins.
		Gels were stained with coomassie brilliant blue, washed with destain and visualised by UVP image analysis.
	www.nibsc.ac.uk /cjd/ds_NP0R00121/coomassie.html (84 words)

	Olympus MIC-D: Polarized Light Gallery
		Brilliant Blue (Coomassie Blue) - Known also as coomassie blue, brilliant blue is a synthetic heterocyclic organic stain, which binds nonspecifically to virtually all proteins.
		The primary function of vitamin K in the body is to assist in normal blood clotting, but it also plays a role in bone calcification.
		Methylene Blue - A chemically basic dye, methylene blue is also known as tetramethyl thionin, and there are various grades used, including methylene blue chloride, which is preferred for biological work.
	www.olympusmicro.com /micd/galleries/polarized (3667 words)

	Staining (biology) - Wikipedia, the free encyclopedia
		Coomassie blue (also brilliant blue) nonspecifically stains proteins a strong blue colour.
		Methylene blue is used to stain animal cells, such as human cheek cells, to make their nuclei more observable.
		Nile blue (or Nile blue A) stains nuclei blue.
	en.wikipedia.org /wiki/Staining_(biology) (2120 words)

	Edman Sequencing Info
		Coomassie blue : 0.5-0.1% Coomassie blue in 50% methanol/10% acetic acid, 5 min.
		We prefer that one of the previous three stains be used first and Coomassie blue used as a last resort.
		One exception is that gel samples should be stained with Coomassie Blue G-250.
	pdtc.rockefeller.edu /Edman/Edmaninfo.html (3095 words)

	TEACHER GUIDE: Protein Fingerprinting
		The bromophenol blue adds color to the solution so students can see the samples as they load them into the gel and determine when the gel has run far enough (the dye is 6-8 cm from the wells).
		They will not be able to see any proteins (bromophenol blue doesn't stain proteins) until they stain the gels with Coomassie blue, a protein dye.
		The sample buffer includes bromophenol blue (allows you to see the sample while you're loading it), glycerol (which is dense and helps the samples sink to the bottom of the wells), SDS (a detergent than denatures and negatively charges proteins so they can move freely through the agarose during electrophoresis), and Tris (a pH buffer).
	biotech.biology.arizona.edu /labs/pro_teach.html (2531 words)

	Computer data acquisition exercise
		Coomassie Blue exists in two color forms, red and blue.
		Upon binding protein, the red form is converted to the blue form.
		Bradford dye solution (10 mg of Serva Blue G from Serva Feinbiochemica, 10 ml of 88% phosphoric acid, 4.7 ml of absolute ethanol, diluted up to 100 ml with distilled water).
	www.siumed.edu /~bkinney/bradford.html (565 words)

	coomassie blue stained gel - BioForum
		I would like to know if a gel after being stained by coomassie blue, and then destained acoordingly, could be used for transfer onto the membrane.
		This was done by comparing the coomassie blue stain with the ponceau stain pattern.
		The thing is that coomassie and destain contains MeOH and acetic acid which fix the proteins, precipitate them in the gel.
	www.protocol-online.org /forums/index.php?showtopic=5627 (374 words)

	Reagents and Buffers - Protein Molecular Weight Markers (Site not responding. Last check: 2007-10-11)
		Prestained markers are ideal to assess the effeciency of protein transfer from the gel to the membrane.These markers are supplied as low and high range.
		After running the gel these markers can be visualized by Coomassie blue stains or by any other conventional stains.
		Since the concentration of Coomassie stain used for staining the gels is lower than that of basic protocol, the background of the gels stays relatively clear.
	www.reagents-and-buffers.com /products/RF_Protein_Analysis/3.htm (1219 words)

	Tp-Blue™ Total Protein Assay
		In the presence of soluble protein the reagent dye forms a blue coloured dye-protein complex, that has an absorbance peak at 595 nm. The large bathochromic shift produced by the dye-protein interaction provides a very effective, ready-to-use, single reagent assay system for soluble proteins.
		Previous Coomassie Brilliant Blue G protein assays were based on the Bradford procedure and suffered from premature precipitation of the dye-protein complex due to the presence of ethanol, (or methanol, as used in commercial products). Alcohol was required for the binding of the dye to the protein.
		Assay solutions had to be measured within a few minutes of mixing, before the protein molecules were dye saturated. This instability produces a non-linear absorbance to protein standard curve.
	www.biocolor.co.uk /proteinblue.asp (417 words)

	W.M. Keck Facility at Yale - MS/MS Protein Identification
		However, Coomassie Blue staining is preferred over the silver staining.
		The staining preference is based on a large 2D gel experiment done in our laboratory where the same proteins (~100 spots total) were digested and identified from a Coomassie Blue stained 2D gel and from a digestion compatible silver stained gel.
		However, in the other silver stained spots, only 10-15% of the peptides observed in the Coomassie blue stained sample were recovered.
	info.med.yale.edu /wmkeck/prochem/msms.htm (569 words)

	Regeneration of Destaining Solution for Electrophoretic Gels
		Following the staining procedure, the gel is typically destained by soaking it in a 20 percent methanol/10 percent acetic acid solution.
		Following the destaining procedure, the destaining solution (methanol/acetic acid) is adulterated with Coomassie blue dye and is not suitable for reuse.
		However, by pouring the destaining solution through a charcoal filter, the blue dye binds to the charcoal, resulting in a purified destaining solution suitable for reuse.
	www.p2000.umich.edu /chemical_waste/cw4.htm (425 words)

	Proteomics FAQs Page
		In general, ~300 protein features can be detected on a mini gel stained with Coomassie blue, while ~1,000 can be detected on a Criterion gel stained with SYPRO ruby.
		For global analysis of protein mixtures, 25-100 µg of protein is sufficient to produce a 2D PAGE pattern detectable by Coomassie brilliant blue using mini gels.
		For individual proteins, the limits of detection are 50-100 ng for colloidal Coomassie brilliant blue or 1-2 ng silver stain.
	dir.niehs.nih.gov /proteomics/faqs.htm (1762 words)

	Biology A/B: Student Instructions (Site not responding. Last check: 2007-10-11)
		If the blue dye of the standards is within 5 millimeters of the end of the gel, stop the electrophoresis by pushing the stop button on the power supply or turn off your power supply.
		If you are the first to finish, you will need to pour the coomassie blue staining solution onto your gel so that it is fully covered.
		If your gel is still too blue, continue the destaining process, making sure you allow the gel and the solution to come to equilibrium, until you get nice clean standards.
	www.accessexcellence.org /AE/SH/NABT_MN/taylor_studentserum.html (1714 words)

	Coomassie Brilliant Blue G-250, 10 g from Bio-Rad - Biocompare Buyer's Guide
		Coomassie Brilliant Blue G-250, 10 g from Bio-Rad
		Coomassie Blue G-250 is a dye for nonspecific staining and detection of proteins.
		Coomassie Blue G-250 must be solubilized in a very acidic environment.
	www.biocompare.com /itemdetails.asp?itemid=4539 (95 words)

	Binding Protein to Coomassie Blue
		Coomassie blue binds to proteins forming a blue color.
		As more protein is added, the solution becomes a deeper blue color.
		The more protein that is added, the deeper the blue color.
	jchemed.chem.wisc.edu /JCESoft/CCA/CCA5/MAIN/2BIOCHEM/BIOCHEM1/COOMASSIE/THUMBS.HTM (86 words)

	OHSU Proteomics Shared Resource: Gel Protocols - Coomassie Stain (Site not responding. Last check: 2007-10-11)
		In general, coomassie stained protein bands offer an excellent chance of success in obtaining useful proteomic information.
		Almost any protein that can be visualized using coomassie will yield interpretable mass spectrometric data.
		Your choice of commercial coomassie R250 stain or 0.1% Coomassie Brilliant Blue R-250 in 50% methanol, 10% acetic acid.
	www.ohsu.edu /proteomics/sampleprep/coomassie.cfm (258 words)

	Electrophoretic analysis of serum proteins in prostate cancer
		In this study a qualitative analysis of Coomassie Brilliant Blue stained proteins separated by SDS-PAGE was undertaken to detect some novel proteins, which could possibly serve as markers for cancer detection.
		Staining solution: 0.5g Coomassie Brilliant Blue R-250 in 450ml methanol, 90ml glacial acetic acid was dissolved in 450ml distilled water.
		Silver staining, which is more sensitive than Coomassie Blue, would tell whether minute quantities of these protein fractions were present.
	www.pmrc.org.pk /cap.htm (2022 words)

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