5-bromo-4-chloro-3-indoxyl phosphate

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Structural formula
Structural formula of 5-bromo-4-chloro-3-indolyl phosphate
General
Surname 5-bromo-4-chloro-3-indoxyl phosphate
other names
  • (5-bromo-4-chloro-1 H -indol-3-yl) dihydrogen phosphate
  • BCIP
  • X-phosphate
  • X-Phos
Molecular formula C 8 H 8 BrClNO 4 P
Brief description

light gray solid (disodium salt)

External identifiers / databases
CAS number
  • 38404-93-2
  • 102185-33-1 (di-sodium salt)
  • 6578-06-9 ( p -toluidine salt)
PubChem 65409
ChemSpider 58873
Wikidata Q238472
properties
Molar mass 326.47 g mol −1
solubility

little in water (20 mg ml −1 , disodium salt)

safety instructions
GHS labeling of hazardous substances
no GHS pictograms
H and P phrases H: no H-phrases
P: no P-phrases
As far as possible and customary, SI units are used. Unless otherwise noted, the data given apply to standard conditions .

5-bromo-4-chloro-3-indoxyl phosphate ( BCIP ) is an artificial, chromogenic substrate for the enzyme alkaline phosphatase (AP).

properties

The enzyme alkaline phosphatase hydrolyzes indolyl phosphate to 5-bromo-4-chloro indolyl and phosphate . The 5-bromo-4-chloroindolyl is oxidized by the oxygen in the air to the deep blue dye 5,5'-dibromo-4,4'-dichloro indigo .

Enzymatic cleavage of X-Phos (BCIP) by means of alkaline phosphatase ( AP ) leads to the formation of 5-bromo-4-chloro-indolyl ( 1 ). After oxidation in air and dimerization, this forms the blue dye 5,5'-dibromo-4,4'-dichloro-indigo ( 2 ).

Since BCIP is sparingly soluble in many solvents, either the disodium salt (soluble in water) or, more often, the p - toluidine salt (soluble in DMF ) is used.

use

In biochemistry , BCIP is used in combination with NBT for the sensitive detection of the enzyme alkaline phosphatase (AP) in the course of immunostaining .

See also

Individual evidence

  1. ↑ a b c Data sheet 5-Bromo-4-chloro-3-indolyl phosphate disodium salt from Sigma-Aldrich , accessed on December 19, 2018 ( PDF ).
  2. ↑ External identifiers or database links for disodium 5-bromo-4-chloro-3- indoxyl phosphate: CAS number: 102185-33-1, EC number: 600-286-2, ECHA InfoCard: 100.106. 217 , PubChem : 6097197 , ChemSpider : 2006838 , Wikidata : Q72452877 .
  3. ↑ External identifiers of or database links to 5-bromo-4-chloro-3-indoxylphosphate-di-p-toluidine salt: CAS number: 6578-06-9, EC number: 229-506-1, ECHA - InfoCard: 100.026.824 , PubChem : 68768006 , ChemSpider : 73132 , Wikidata : Q72452880 .
  4. ↑ James McGadey: A tetrazolium method for non-specific alkaline phosphatase , in: Histochemie , 1970 , 23 , pp. 180-184; PMID 5507043 ; doi : 10.1007 / BF00305851 .
  5. ↑ David A. Knecht, Randall L. Dimond: Visualization of antigenic proteins on Western blots , in: Anal. Biochem. , 1984 , 136 , pp. 180-184; PMID 6201085 ; doi : 10.1016 / 0003-2697 (84) 90321-X .