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2-Nitrophenyl-β-D-Galactopyranoside BioChemica

Assay (HPLC): min. 99 %
Code
A1272
CAS
369-07-3
Molecular Formula
C12H15NO8
Molar mass
301.26 g/mol

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Box prices only valid with purchase of full box.

code packaging size price per unit box price per unit
Code & packaging Price per piece
A1272,0005
code
A1272,0005
packaging size
5 g
price per unit
single 62,50€
box price per unit
A1272,0025
code
A1272,0025
packaging size
25 g
price per unit
single 232,80€
box price per unit
A1272,0200
code
A1272,0200
packaging size
200 g
price per unit box price per unit
Physical Description:
Solid
Product Code:
A1272
Product Name:
2-Nitrophenyl-β-D-Galactopyranoside BioChemica
Specifications:
Assay (HPLC): min. 99 %
α20°C/D; 1 %, H2O: -67° - -71°
2-Nitrophenol: max. 0.03 %
WGK:
1
Storage:
-20°C
protected from light
EINECS:
206-716-1
CS:
29389090
Download TDS file for complete specifications

Comments

ONPG is one of the major substrates in β-galactosidase enzyme assays. The properties of this enzyme have been studied in detail (e. g. ref. 1-3). Expression plasmids with the coding sequence of β-Galactosidase are widely used as internal controls for transfection experiments in expression and transcription studies, usually under the control of a strong enhancer (SV40, CMV, RSV, HIV etc.). The enzyme activity is easily determined from cell extracts (e. g. ref. 4). The cleavage of ONPG releases the yellow dye 4-nitrophenyl and absorption is measured at 405 - 420 nm by spectrophotometry. A commonly used reaction buffer is\: 60 mM Na₂HPO₄ / 39 mM NaH₂PO₄, pH 7.0 / 10 mM KCl / 1 mM MgSO₄ / 2 mM DTT and 1 mg/ml ONPG. ONPG is of low solubility. The solid ONPG is added to the reaction buffer and dissolved by warming the buffer to 37°C and/or well mixing. DTT is added freshly (like ONPG) from a 1 M stock solution (4).Note\: For the investigations mentioned above, the bacterial β-galactosidase (E. coli) is used. The pH-optimum of this enzyme is pH 7.3. In mammalian cells exsists a lysosomal β-galactosidase with a pH-optimum at 3.5. Make sure, that the pH of the reaction buffer is correct, otherwise there could be a significant background by the endogenous enzyme!

Literature

(1)Wallenfels, K. et al. (1960) Biochem. Z. 333, 377-394Untersuchungen über milchzuckerspaltende Enzyme. (2)Wallenfels, K. (1962) Methods Enzymol. 5, 212-219β-Galactosidase\: Assay methods. (3)Levvy, G.A. & Conchie, J. (1966) Methods Enzymol. 8, 571-584Mammalian glycosidases and their inhibition by Aldonolactones. (4)Janknecht, R. et al. (1995) Oncogene 10, 1209-1216SAP1a is a nuclear target of signaling cascades involving ERKs.