Coelenterazine, 1 mg
Molar mass (M) 423,48 g/mol
Melting point (mp) 178 °C
Storage temp. -20 °C
Transport temp. ambient temp.
CAS No. [55779-48-1]
excl. VAT. | 1 mg per Pack Qty.
Art. No. 4094.3
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Coelenterazine ≥95 %, for biochemistry
Substrate of the Renilla luciferase used for monitoring of the mitochondrial Calcium flux and of reportergenes. Coelenterazine is applied in bioluminescence resonance energy transfer (BRET) assays, ELISAs, HTS and in chemiluminescent detection of superoxide anion and peroxynitrite in cells or tissues.
|Signal product||soluble (ELISA, BRET)|
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Stock solutions (All solutions are stable for at least 1 year at 4 °C):
0.5 g NBT in 10 ml 70 % dimethylformamide;
0.5 g BCIP (p-toluidine salt) in 10 ml 100 % dimethylformamide.
Incubation buffer for alkaline phosphatase:
100 mM NaCl, 5 mM MgCl2,100 mM Tris (pH 9.5).
Fresh substrate solution: 66 μl NBT stock solution + 10 ml incubation buffer, mix well, add 33 μl BCIP stock solution. Use within 1 hour.
Blot development: Approx. 10 ml substrate solution per 15 x 15 cm2 membrane surface. Develop at room temperature until bands become visible (approx. 30 mins).
Reaction stop: Rinse with PBS/20 mM EDTA.
Dissolve 1 mg TMB in 0.1 ml dimethylsulfoxide (Art. No. 4720); add 9.9 ml of a 0.1 M sodium acetate solution (pH 6.0) (Powder: Art. No. 6779), filter and add H2O2 (Art. No. 8070) (final concentration 0.01 %).
Always prepare freshly!
Incubation 10-30 mins at room temperature (approx. 50 μl per microtitre well; finally add 50 μl 1 M H2SO4, Art. No. X873, per well).
Photometric quantitation at 450 nm.
Reference: Bos, E.S. et al., (1981) J. Immunoassay 2, (3/4), 187.
|Assay (TLC)||≥95 %|