Bromophenol blue (3′,3″,5′,5″-tetrabromophenolsulfonphthalein, BPB), albutest is used as a

pH indicator A pH indicator is a halochromism, halochromic chemical compound added in small amounts to a Solution (chemistry), solution so the pH (acidity or Base (chemistry), basicity) of the solution can be determined visually or spectroscopically by chang ...

, an

electrophoretic color marker An electrophoretic color marker is a chemical used to monitor the progress of agarose gel electrophoresis and polyacrylamide gel electrophoresis (PAGE) since DNA, RNA, and most proteins are colourless. The color markers are made up of a mixture of ...

, and a dye. It can be prepared by slowly adding excess bromine to a hot solution of

phenolsulfonphthalein Phenol red (also known as phenolsulfonphthalein or PSP) is a pH indicator frequently used in cell biology laboratories. Chemical structure and properties Phenol red exists as a red crystal that is stable in air. Its solubility is 0.77 grams per ...

in glacial acetic acid.

Acid–base indicator

As an acid–base indicator, its useful range lies between pH 3.0 and 4.6. It changes from yellow at pH 3.0 to blue at pH 4.6; this reaction is reversible. Bromophenol blue is structurally related to

phenolphthalein Phenolphthalein ( ) is a chemical compound with the chemical formula, formula carbon, C20hydrogen, H14oxygen, O4 and is often written as "HIn", "HPh", "phph" or simply "Ph" in shorthand notation. Phenolphthalein is often used as an indicator in ...

(a popular indicator).

Color marker

Bromophenol is also used as a colour marker to monitor the process of

agarose gel electrophoresis Agarose gel electrophoresis is a method of gel electrophoresis used in biochemistry, molecular biology, genetics, and clinical chemistry to separate a mixed population of macromolecules such as DNA or proteins in a matrix of agarose, one of the t ...

and

polyacrylamide gel electrophoresis Polyacrylamide gel electrophoresis (PAGE) is a technique widely used in biochemistry, forensic chemistry, genetics, molecular biology and biotechnology to separate biological macromolecules, usually proteins or nucleic acids, according to their ...

. Since bromophenol blue carries a slight negative charge at moderate pH, it will migrate in the same direction as

DNA Deoxyribonucleic acid (; DNA) is a polymer composed of two polynucleotide chains that coil around each other to form a double helix. The polymer carries genetic instructions for the development, functioning, growth and reproduction of al ...

protein Proteins are large biomolecules and macromolecules that comprise one or more long chains of amino acid residue (biochemistry), residues. Proteins perform a vast array of functions within organisms, including Enzyme catalysis, catalysing metab ...

in a gel; the rate at which it migrates varies according to gel density and

buffer Buffer may refer to: Science * Buffer gas, an inert or nonflammable gas * Buffer solution, a solution used to prevent changes in pH * Lysis buffer, in cell biology * Metal ion buffer * Mineral redox buffer, in geology Technology and engineeri ...

composition, but in a typical 1%

agarose Agarose is a heteropolysaccharide, generally extracted from certain red algae. It is a linear polymer made up of the repeating unit of agarobiose, which is a disaccharide made up of D-galactose and 3,6-anhydro-L-galactopyranose. Agarose is on ...

gel in a 1X

TAE buffer TAE buffer is a buffer solution containing a mixture of Tris base, acetic acid and EDTA. In molecular biology, it is used in agarose electrophoresis typically for the separation of nucleic acids such as DNA and RNA. It is made up of Tris-acetate ...

TBE buffer TBE or Tris/Borate/EDTA, is a buffer solution containing a mixture of Tris base, boric acid and EDTA. In molecular biology, TBE and TAE buffers are often used in procedures involving nucleic acids, the most common being electrophoresis. Tris-aci ...

, bromophenol blue migrates at the same rate as a DNA fragment of about 300

base pair A base pair (bp) is a fundamental unit of double-stranded nucleic acids consisting of two nucleobases bound to each other by hydrogen bonds. They form the building blocks of the DNA double helix and contribute to the folded structure of both DNA ...

s, in 2% agarose as 150 bp.

Xylene cyanol Xylene cyanol can be used as an electrophoretic color marker, or tracking dye, to monitor the process of agarose gel electrophoresis and polyacrylamide gel electrophoresis. Bromophenol blue and orange G can also be used for this purpose. Once mi ...

and

orange G Orange G also called C.I. 16230, Acid Orange 10, or orange gelb is a synthetic azo dye used in histology in many staining formulations. It usually comes as a disodium salt. It has the appearance of orange crystals or powder. Staining Orange G ...

may also be used for this purpose.

Dye

Bromophenol blue is also used as a dye. At neutral pH, the dye absorbs red light most strongly and transmits blue light. (Its peak absorbance is 590 nm at a basic pH of 12.) Solutions of the dye, therefore, are blue. At low pH, the dye absorbs ultraviolet and blue light most strongly and appears yellow in solution. In solution at pH 3.6 (in the middle of the transition range of this pH indicator) obtained by dissolution in water without any pH adjustment, bromophenol blue has a characteristic green red colour, where the apparent colour shifts depending on the concentration and/or path length through which the solution is observed. This phenomenon is called dichromatic color. Bromophenol blue is the substance with the highest known value of

Kreft's dichromaticity index Kreft's dichromaticity index (DI) is a measure for quantification of dichromatism. It is defined as the difference in hue angle (Δhab) between the color of the sample at the dilution, where the chroma (color saturation) is maximal, and the color ...

. This means it has the largest change in colour

hue In color theory, hue is one of the properties (called color appearance parameters) of a color, defined in the CIECAM02 model as "the degree to which a stimulus can be described as similar to or different from stimuli that are described as ...

, when the thickness or concentration of observed sample increases or decreases.

References

External links

Bromophenol blue
at OpenWetWare {{DEFAULTSORT:Bromophenol Blue PH indicators Triarylmethane dyes Phenol dyes Bromobenzene derivatives Benzoxathioles

Acid–base indicator

Color marker

Dye

See also

References

External links