Fluorescent Dopamine | Fluorescent Neurotransmitter - Dansyl 5-C Conjugated Dopamine | FIVEphoton Biochemicals | Dansyl-Dopamine | Part DnsylD-1

Fluorescent Dopamine | Fluorescent Neurotransmitter - Dansyl 5-C Conjugated Dopamine | FIVEphoton Biochemicals | Dansyl-Dopamine | Part DnsylD-1
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Price: $308.00
Availability: In Stock
Model: DnsylD-1
Manufacturer: Fivephoton Biochemicals

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Fluorescent Dopamine Neurotransmitter 

(Dopamine labeled with dansyl attached through a 5-carbon spacer arm:  Ex/Em: 333/515 nm).  Syn:  Dansyl-Dopamine, Part DnsylD-1TM
 
Updated 9-2022
Proprietary product of Fivephoton Biochemicals

Icon Image:  A low magnification view of multiple labeled cells over a well of a 96-well culture dish.

Highlights
  1. Pharmalogical properties, including affinity, virtually identical to unconjugated dopamine.
  2. DnsylD-1 operates selectively in uptake and binding assays at nanomolar concentrations. 
  3. The native dopamine chemical structure is the major component of this fluorescent catecholamine, generating a fluorescent neurotransmitter molecule with high affinity toward all dopamine receptor subtypes as well as to the dopamine transporter.
  4. The small dansyl tag with a 5-carbon polylinker to dopamine generates a stable, bright fluorescent signal with negligible steric hindrance. 
  5. The dansyl fluorescent tag offers a large stokes shift that minimizes autofluorescence and non-specific background signals:  (Ex/Em:  313/515nm).
  6. The fluorescent signal of DnsylD-1TM is blockable by standard inhibitors to dopamine receptors and DAT.
  7. Standard inhibitors can be employed to distinguish the interaction of DnsylD-1TM with dopamine receptors and DAT.
  8. Fluorescence from DnsylD-1TM colocalizes with tyrosine hydroxylase and identifies dopaminergic neurons.
  9. To assay for dopamine receptor binding, first suspend the provided solid in DMSO (10 - 50 µl).  Centrifuge to clarify, and collect the DMSO soluble fraction, which can be aliquoted and stored at -20oC:  DnsylD-1TM when suspended in DMSO and frozen at -20oC is stable for at least 2 years.  Dilute the DMSO solubilized material into your assay buffer (for example PBS-BSA) to the desired assay concentration.  Detect DnsylD-1TM by fluorescent microscopy, with a fluorescent plate reader or by flow cytometry.  Set the fluorescent detection device to Ex/Em:  333/515 nm.

Quantities1
 Mass
Moles
Cost
0.5mg
1 µmole $308
1mg 2 µmole $523.8
5mg
10 µmole $1512
15mg
30 µmole $4200

Bulk quantities and custom synthesis are available.  Please inquire with customersupport@fivephoton.com

1To estimate the amount of DnsylD-1TM required for your needs, use an assay concentration 10 to 100 fold the Kd of dopamine for the dopamine receptor subtype or transporter that you are investigating (use a literature review to identify the average reported Kd value of the dopamine receptor subtype or transporter).  Then estimate the total external assay buffer volume that is required for your assay (for example, 50 µl of assay buffer per well in a 96 well dish multiplied by the number of wells) to calculate the amount of DnsylD-1TM required for the entire assay.  For technical assistance, contact technicalsupport@fivephoton.com or by telephone: 800.462.4507 (International:  858.395.4026).


Specifications
  • Spectral characteristics and assay wavelengths:  Ex:Em:  333:515 nm
  • MW:  499.62 (Mass and QC verified by MALDI)
  • Appearance:  lyophilized off-white solid
  • Purity:  ≥ 95% (HPLC; view COA for lot purity)
  • Solubility:  DMSO
  • Analytical QC Test:  MALDI MS and HPLC
  • Use at 10X Kd of unlabeled dopamine toward respective dopamine receptor subtype or transporter
  • Storage (lyophilized powder):  4oC, dark, desiccate.  Storage when solubilized in DMSO:  -20oC, aliquot.
  • Shipping:  Ambient temperature.

 
 
Specification Sheet

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Fluorescent Dopamine Neurotransmitter Ligand (Dansyl-Dopamine), Overview
This fluorescent labeled catecholamine neurotransmitter analog of dopamine (Product Name:  DnsylD-1TM), with dansyl attached by a 5-carbon spacer arm to dopamine, provides a unique pharmacological tool.  DnsylD-1TM is a stable, receptor selective and high affinity pharmacological ligand that can be detected by fluorescent microscopy and FLIPR devices.  Unlike recently reported synthesized fluorescent neurotransmitter analogs, DnsylD-1TM contains the entire native dopamine structural moiety attached with the 5-carbon polylinker to the small fluorescence label dansyl.  This structural modification minimizes steric hindrance, making DnsylD-1TM an effective reagent for dopamine ligand binding and transporter uptake assays at namomolar concentrations. 

The dansyl fluorescent tag offers a large stokes shift between excitation and emission maxima (Ex:Em, 313:515 nm) facilitating high signal to noise detection with minimal autofluorescence, unlike other fluorescent tags such as FITC.  The dansyl moiety also changes spectral characteristics on intermolecular contact, facilitating biophysical characterization of ligand-receptor interaction.  DnsylD-1TM is a suitable substitute for radioactive ligand.

Applications for Fluorescent Dopamine Ligand-Neurotransmitter (DnsylD-1TM)
DnsylD-1TM has the following applicability due to the small molecular size of the dansyl fluorescent label, a long spacer arm between the dopamine structure and the fluorescent tag, stability in aqueous solution and low autofluorescence of the surrounding cellular or tissue medium.
  1. Pharmacological applications as a substitute for radioactive ligand.
  2. Dopaminergic neuron detection:  Biomarker detection.
  3. Dopamine uptake analysis.
  4. Histochemical imaging and quantitation of dopamine receptor expression in vivo and vitro.
  5. Dopamine metabolism and turnover analysis.
  6. Analysis of disease conditions associated with altered dopamine synthesis, dopamine receptor expression or dopamine uptake:  human disease research applicability in the following:  a)  Parkinson's disease.  b)  Psychological disorder, including ADHD, schizophrenia and substance abuse.  c)  Physiological regulation of cardiovascular, renal and immune function.

Images:  DnsylD-1TM (Dansyl-Dopamine) bound to neuroblastoma cells and observed by low magnification fluorescent microscopy over a well of a 96-well plate.  Multiple cells are displayed.  

Top Panel.  Rat neuroblastoma cells, which endogenously express dopamine receptors, grown in a well of a 96-well dish incubated for 30 min with DnsylD-1TM at a concentration 10-fold the Kd concentration for dopamine to D2 dopamine receptors.  The image was taken at 50X magnification on top of a well with a fluorescent microscope.

Bottom Panel.  Pre-exposure of 1000 X Kd concentration of unlabeled dopamine for 15min prior to exposure of DnsylD-1TM at 10X the Kd concentration for D2 receptors, as in the top image.  Pre-exposure of unlabeled dopamine effectively blocks binding of DnsylD-1TM demonstrating high affinity and selectivity.


Product Reference
May S, Andreasson-Ochsner M, Fu Z, Low YX, Tan D, de Hoog HP, Ritz S, Nallani M, Sinner EK.  2013. In Vitro Expressed GPCR Inserted in Polymersome Membranes for Ligand-Binding Studies.  Angew. Chem. Int. Ed, 52, 749 –753.  Link to article
 
Methods description for dopamine receptors and transporter. 2019.  Link
 

* For research use only.  Not for diagnostics or therapeutics.   

Safety:   Irritant.  Avoid ingestion, skin and eye contact and inhalation.  Not a drug.  For research use only.
Storage:  Provided as a lyophilized solid. Store at  4oC;  minimum 2 years:  -20oC longer term storage.  Desiccate.  Can also be solubilized in DMSO and stored at -20oC for a minimum of 2 years.  Limit exposure to light.
Shipping Options:  Shipped at ambient temperature.  2-day and overnight delivery within North American (United States and Canada).  Please inquire with customersupport@fivephoton.com for international delivery costs.  International customers can also provide their Fedex or DHL accounts to cover shipping. 
 
Ordering.  DnsylD-1TM can be ordered directly from Fivephoton Biochemicals, or from Fisher Scientific (product number: NC0919495 (0.5mg), Cedarlane or Antibodies-Online in the United States.  Ordering is also available from international distributors of Fivephoton Biochemicals.
kw. fluorescent neurotransmitter, dopamine transporter assay, dopamine transporter uptake assay, dopamine receptor biomarker,  dopaminergic neurons fluorescent detection reagent, dopamine receptor binding assay, DAT, fluorescent catecholamine, vesicular monoamine transporter fluorescent detection. dopamine




 

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Specification Sheet
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COA click here
MSDS click here