Cell Signaling Technology

Product Pathways - Neuroscience

SignalSilence® Dopamine β-Hydroxylase (DBH) siRNA I #7664

Applications Reactivity
Transfection H (M) (Mk)

Reactivity Key:  H=Human  M=Mouse  Mk=Monkey
Species cross-reactivity is determined by western blot. Species enclosed in parentheses are predicted to react based on 100% sequence homology.

Western Blotting

Western Blotting

Western blot analysis of extracts from SH-SY5Y cells, transfected with 100 nM SignalSilence® Control siRNA (Unconjugated) #6568 (-) or SignalSilence® Dopamine β-Hydroxylase (DBH) siRNA I (+), using Dopamine β-Hydroxylase (DBH) Antibody #8586 (upper) or β-Actin (13E5) Rabbit mAb #4970 (lower). The Dopamine β-Hydroxylase (DBH) Antibody confirms silencing of dopamine β-hydroxylase expression, while the β-Actin (13E5) Rabbit mAb is used as a loading control.

Description

SignalSilence® Dopamine β-Hydroxylase (DBH) siRNA I from Cell Signaling Technology (CST) allows the researcher to specifically inhibit dopamine β-hydroxylase expression using RNA interference, a method whereby gene expression can be selectively silenced through the delivery of double stranded RNA molecules into the cell. All SignalSilence® siRNA products from CST are rigorously tested in-house and have been shown to reduce target protein expression by western analysis.

Quality Control

Oligonucleotide synthesis is monitored base by base through trityl analysis to ensure appropriate coupling efficiency. The oligo is subsequently purified by affinity-solid phase extraction. The annealed RNA duplex is further analyzed by mass spectrometry to verify the exact composition of the duplex. Each lot is compared to the previous lot by mass spectrometry to ensure maximum lot-to-lot consistency.

Directions for Use

CST recommends transfection with 100 nM SignalSilence® Dopamine β-Hydroxylase (DBH) siRNA I 48 to 72 hours prior to cell lysis. For transfection procedure, follow protocol provided by the transfection reagent manufacturer. Please feel free to contact CST with any questions on use.

Each vial contains the equivalent of 100 transfections, which corresponds to a final siRNA concentration of 100 nM per transfection in a 24-well plate with a total volume of 300 μl per well.

Background

Dopamine β-Hydroxylase (DBH) is an enzyme of the copper type II ascorbate-dependent mono-oxygenase family. This enzyme forms homotetramers composed of two noncovalently bound disulfide-linked dimers and is found as both membrane-associated and soluble forms (1-3). The soluble form is present in the lumen of secretory granules (4) and is released from cells by exocytosis (5). DBH converts dopamine to noradrenaline (6). Deficiency in this enzyme causes a rare disease characterized by a complete absence of noradrenaline and adrenaline in plasma together with increased plasma dopamine levels (7). Orthostatic hypotension, the main symptom of DBH deficiency, can be alleviated by administration of dihydroxyphenylserine, a synthetic precursor of noradrenaline (8).

  1. Smith, W.J. and Kirshner, N. (1967) Mol Pharmacol 3, 52-62.
  2. Lagercrantz, H. (1976) Neuroscience 1, 81-92.
  3. Winkler, H. (1976) Neuroscience 1, 65-80.
  4. Laduron, P.M. (1975) FEBS Lett 52, 132-4.
  5. Weinshilboum, R.M. et al. (1971) Science 174, 1349-51.
  6. Kaufman, S. and Friedman, S. (1965) Pharmacol Rev 17, 71-100.
  7. Robertson, D. et al. (1986) N Engl J Med 314, 1494-7.
  8. Biaggioni, I. and Robertson, D. (1987) Lancet 2, 1170-2.

Application References

Have you published research involving the use of our products? If so we'd love to hear about it. Please let us know!

Companion Products

Limited Use Label License, RNA interference: This product is licensed under European Patent 1144623 and foreign equivalents from Ribopharma AG, Kulmbach, Germany and is provided only for use in non-commercial research specifically excluding use (a) in drug discovery or drug development, including target identification or target validation, by or on behalf of a commercial entity, (b) for contract research or commercial screening services, (c) for the production or manufacture of siRNA-related products for sale, or (d) for the generation of commercial databases for sale to Third Parties. Information about licenses for these and other commercial uses is available from Ribopharma AG, Fritz-Hornschuch-Str. 9, D-95326 Kulmbach, Germany.

For Research Use Only. Not For Use In Diagnostic Procedures.

Products