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BCIP (5-bromo-4-chloro-3-indolyl phosphate, p-toluidine salt) is a chromogenic substrate for alkaline phosphatase, used in combination with the oxidant NBT (nitro blue tetrazolium) to enhance blue color development.
When alkaline phosphatase conjµgates are used for detection, BCIP is recommended for the following applications: Southern blotting, Western blotting, Northern blotting and dot/slot blots.
It is recommended to prepare a 50mg/mL stock solution in DMF.
Protocol for Preparation of BCIP/NBT Substrate Solution:
Add the following components
- 10mL of the Alkaline Phosphatase (AP) buffer (100mM Tris-HCl (pH9.5), 100mM NaCl and 10mM MgCl2)
- 33µL BCIP (50mg/mL in DMF)
- 44µL NBT (75mg/mL in 70% DMF).
NOTE: Avoid exposure to light - Prepare fresh developing solution and use within an hour - BCIP and NBT solutions in DMF do not freeze. They are stable for approximately 2 years when stored at -20°C in the dark.
Assay: min. 98% (HPLC)
pH (5% in water): 1.0-2.0
Solubility (water, 20°C): 582 g/L
MP = 120°C
Chloride: max. 0.001%
Pb: max. 0.005%
Free acid: max. 0.1%
C8H6BrClNO4P x C7H9N
MW = 433.64 g/mol
Sicherheits Hinweise / SafetyH-Sätze: H315, H319, H335
Klassifizierungen / ClassificationEC-Nr: 229-506-1
Dokumente - Protokolle - Downloads
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