Transglutaminase (serves as a glutaminyl substrate; guinea pig liver transglutaminase) - no specific IC50/Ki values provided for this enzymatic reaction [1]
Transglutaminase (serves as an aminyl substrate) - no specific IC50/Ki values provided [4]
Calmodulin (antagonizes calmodulin-dependent activation of phosphodiesterase) - apparent Ki for calmodulin-stimulated phosphodiesterase: 120 μM [4]
Lipid A (bacterial lipopolysaccharide) - binding with Kd ranging from 16 μM to 26 μM [5]
Core glycolipid (from Salmonella minnesota Re595) - binding with Kd ranging from 22 μM to 28 μM [5]

Dansylcadaverine's capacity to competely inhibit the cross-linking of fibrin molecules and act as a substrate for transglutaminase is demonstrated by its inhibitory activity [2]. A cationic fluorescent probe called dansylcadaverine binds to lipid A and bacterial lipopolysaccharide. Other substances that have an affinity for endotoxins can displace it competitively [3].

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Dansylcadaverine serves as a specific glutaminyl substrate for transglutaminase and can be fluorescently labeled onto thymosin β4. Two glutamine residues (Gln-23 and Gln-36) of thymosin β4 were mainly involved in the transglutaminase reaction, while Gln-39 was derivatized with low efficiency. Labeled derivatives (TG47, TG48, TG51, TG55) were able to inhibit polymerization of G-actin [1].
Dansylcadaverine (100-500 μM) inhibits adhesion of Chinese hamster ovary (CHO) cells to serum-coated substrata in a reversible, dose-dependent manner. The concentration required to produce 50% inhibition of adhesion was approximately 300 μM. At 100 μM, no effect on cell attachment was observed; at 300 μM, marked inhibition of attachment (only 20% of cells attached at 15 min); at 500 μM, complete inhibition of attachment. The compound also induced morphological changes including increased numbers of rounded and detached cells at 500 μM [4].
Dansylcadaverine (50-100 μM) inhibits calmodulin-stimulated brain cyclic nucleotide phosphodiesterase activity in a dose-dependent manner with an apparent Ki of 120 μM, while not inhibiting basal phosphodiesterase activity [4].
Dansylcadaverine binds to lipid A from Salmonella minnesota with an apparent stoichiometry of two equivalent binding sites and Kd of 16.6 × 10⁻⁶ M (by Scatchard analysis) or 26 × 10⁻⁶ M (by Scatchard-type plot). Addition of lipid A to Dansylcadaverine results in marked enhancement of fluorescence intensity accompanied by a blue shift in emission maximum, with steady-state emission polarization values increasing from 0.031 to 0.157 [5].
Dansylcadaverine binds to core glycolipid (CGL) from Salmonella minnesota Re595 with three apparently equivalent binding sites and Kd of 22 × 10⁻⁶ M (by Scatchard analysis) or 28.8 × 10⁻⁶ M (by Scatchard-type plot) [5].
Dansylcadaverine is displaced from lipid A by polymyxin B in a biphasic manner, with the displacement curve suggesting two different classes of binding sites on lipid A for polymyxin, corresponding to Kd values of 0.4 × 10⁻⁶ M and 1.5 × 10⁻⁶ M for high- and low-affinity interactions respectively [5].
Dansylcadaverine is displaced from core glycolipid by polymyxin B with apparent dissociation constants of 1.1 × 10⁻⁶ M and 5.8 × 10⁻⁶ M [5].
Dansylcadaverine (50 μmol/l) is used to label acidic vesicular organelles (autophagic vacuoles) in MCF7 cells. Cells were incubated with MDC at 37°C for 15 min and washed with PBS before observation under fluorescence microscope. CuO NPs (4, 6, 12 μg/ml) induced autophagy in a dose-dependent manner as revealed by MDC staining [2].
Dansylcadaverine staining was used to detect autophagic vacuoles in HNSCC cells (HN12) treated with CYT997 (100 nM for 24 h), revealing dot-like structures in the cytoplasmic and perinuclear regions of drug-treated cells [3].

Transglutaminase activity was measured by the Ca²⁺-dependent incorporation of [³H] Dansylcadaverine (15,000 dpm/pmol) into casein. The reaction mixture contained 20 mM Tris-HCl (pH 7.4), 2 mg/ml N,N-dimethylcasein, 0.8 μM [³H] Dansylcadaverine, 15 mM β-mercaptoethanol and either 5 mM EGTA or 5 mM CaCl₂. All assays were linear with time and protein concentration [4].
Calmodulin-stimulated brain cyclic nucleotide phosphodiesterase activity was assayed by measuring time-dependent conversion of [³H] cGMP (2.5 μM) to [³H] guanosine in the presence of 1 mM CaCl₂, 40 mM Tris-HCl (pH 8.0), 1.5 mM brain calmodulin, 1.25 mg/ml bovine serum albumin, and various concentrations of Dansylcadaverine (0-500 μM) [4].
For binding studies, Dansylcadaverine binding parameters to lipid A and core glycolipid were determined by fluorimetric titration in 100 mM phosphate-buffered saline (pH 7.4). Excitation wavelength was 340 nm. Scatchard analyses were performed, requiring nonlinear extrapolation of Fₘ (fluorescence yield of bound probe at unit probe concentration) using a quadratic function modelled on data points with a Simplex function minimizing algorithm due to nonlinear double-reciprocal plots [5].
Displacement experiments using Dansylcadaverine were performed with a 10-12 molar excess of probe to ensure a high bound/free ratio of lipid A or CGL. Occupancy was calculated as (F - F₀)/(Fₘₐₓ - F₀), where F₀ is fluorescence intensity of Dansylcadaverine alone, Fₘₐₓ is intensity in presence of lipid A, and F are intensities at different displacer concentrations. The Horovitz-Levitzki method was used for analyzing displacement data [5].
Transglutamination of thymosin β₄ with Dansylcadaverine was carried out by incubating 120 μM thymosin β₄ with 5 mM Dansylcadaverine in buffer consisting of 10 mM Tris-HCl (pH 7.4), 15 mM CaCl₂, 3 mM DTT. The reaction was started by addition of 0.1 U transglutaminase. At indicated times, aliquots were diluted in 0.1% TFA to stop the reaction and analyzed by HPLC [1].

For adhesion assays, CHO cells metabolically labeled with [³H] leucine were preincubated for 30 min with Dansylcadaverine (100-500 μM) and then assayed for attachment to plastic tissue culture dishes in continued presence of the drug. Attachment was measured at 15, 30, 60, and 90 min. Cell viability was measured by trypan blue exclusion [4].
For autophagy detection, MCF7 cells (2.4 × 10⁴) were seeded into 35 mm plates. After 24 h incubation, CuO NPs were added with increasing concentrations (4, 6, 12 μg/ml) in the presence or absence of 3-MA (autophagy inhibitor) for different time periods. Cells were then incubated with 50 mmol/l Dansylcadaverine (MDC) at 37°C for 15 min and washed with 1× PBS three times with 5 min intervals. Cells were observed under a fluorescence microscope [2].
For autophagic flux measurement, MCF7 cells were transfected with pBABE-puro mCherry-EGFP-LC3B plasmid for 16 h then treated with CuO NPs at 12 μg/ml for different times with or without inhibitor. After treatment, cells were washed, mounted on glass slides, and monitored for EGFP and m-Cherry under a fluorescence microscope [2].
For HNSCC cell studies, HN12 cells were treated with 100 nM CYT997 for 24 h in the presence or absence of 20 μM hydroxychloroquine (HCQ). Autophagy was determined by Dansylcadaverine staining, and cells were examined under fluorescence microscopy to detect dot-like structures in cytoplasmic and perinuclear regions [3].
For cell viability assays in HNSCC cells, cells were treated with Dansylcadaverine (as part of autophagy detection) in combination with CYT997. Cell viability was determined by MTS assays and flow cytometry with an amine-reactive fluorescent dye Zombie Aqua™. Apoptosis was determined by flow cytometry with Annexin V-FITC staining and Western blot with antibodies against cleaved PARP [3].

Dansylcadaverine at concentrations up to 500 μM for 30 min preincubation did not affect CHO cell viability as measured by trypan blue exclusion (viability >95% at all concentrations tested) [4].
Dansylcadaverine treatment (500 μM for 30 min) effects on CHO cells were reversible; after 20-fold dilution of the drug, cells attached, spread, and attained morphology indistinguishable from untreated cells [4].

[1]. Thymosin beta(4) serves as a glutaminyl substrate of transglutaminase. Labeling with fluorescentdansylcadaverine does not abolish interaction with G-actin. FEBS Lett. 1999 Dec 24;464(1-2):14-20.

[2]. Interplay between autophagy and apoptosis mediated by copper oxide nanoparticles in human breast cancer cells MCF7. Biochim Biophys Acta. 2014 Jan;1840(1):1-9.

[3]. Autophagy blockade sensitizes human head and neck squamous cell carcinoma towards CYT997 through enhancing excessively high reactive oxygen species-induced apoptosis. J Mol Med (Berl). 2018;96(9):929-938.

[4]. Inhibition of the adhesion of Chinese hamster ovary cells by the naphthylsulfonamides dansylcadaverine and N-(6-aminohexyl)-5-chloro-1-naphthylenesulfonamide (W7). Biochim Biophys Acta. 1983;762(3):414-419.

[5]. Analysis of the binding of polymyxin B to endotoxic lipid A and core glycolipid using a fluorescent displacement probe. Biochim Biophys Acta. 1992;1165(2):147-152.

Monodanthiocarbamate is a sulfonamide formed by the condensation of the sulfonic acid group of 5-(dimethylamino)naphthalene-1-sulfonic acid with an amino group of cadaverine. It can be used as a fluorescent dye, a protective agent, and an EC 2.3.2.13 (protein-glutamine γ-glutamyl transferase) inhibitor. It is a sulfonamide, aminonaphthalene, tertiary amine, and primary amine compound. Its function is related to that of cadaverine.

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Dansylcadaverine was originally developed as a specific inhibitor of transglutaminase-mediated fibrin crosslinking. Its inhibitory activity reflects its ability to serve as a substrate for transglutaminases and to block competitively the crosslinking of fibrin molecules [4].
Dansylcadaverine has been reported to be a potent inhibitor of receptor-mediated endocytosis of diverse molecules including α₂-macroglobulin, epidermal growth factor, catecholamines, insulin, triiodothyronine, and vesicular stomatitis virus [4].
Dansylcadaverine and the structurally related compound W7 (N-(6-aminohexyl)-5-chloro-1-naphthylenesulfonamide) are both naphthylsulfonamides that can inhibit calmodulin-dependent processes. W7 is approximately 4-times more potent than Dansylcadaverine in antagonizing calmodulin (apparent Ki 30 μM vs 120 μM) [4].
Dansylcadaverine serves as a glutaminyl substrate for transglutaminase at concentrations present inside cells (e.g., in human platelets at about 500 μM). It is likely that thymosin β₄ serves as a glutaminyl substrate for factor XIIIa in vivo [1].
Dansylcadaverine can be used as a displacement probe to quantitate interactions of various substances with lipid A and endotoxins, offering a convenient method for screening compounds for lipid A-antagonistic properties [5].
Dansylcadaverine (monodansylcadaverine, DNC) was synthesized from n-5-aminopentyl-5-dimethylamino-1-naphthalene sulfonamide. The compound was verified to be homogeneous by TLC on silica-gel using petroleum ether/chloroform/methanol (2:2:1). 270 MHz ¹H NMR, IR and UV spectra confirmed the structure [5].

C17H25N3O2S

335.4643

335.167

10121-91-2

4247

White to off-white solid powder

1.19g/cm3

505.5ºC at 760mmHg

137-140ºC

259.5ºC

9.54E-11mmHg at 25°C

1.594

4.485

2

5

8

23

447

0

MLEBFEHOJICQQS-UHFFFAOYSA-N

InChI=1S/C17H25N3O2S/c1-20(2)16-10-6-9-15-14(16)8-7-11-17(15)23(21,22)19-13-5-3-4-12-18/h6-11,19H,3-5,12-13,18H2,1-2H3

N-(5-aminopentyl)-5-(dimethylamino)naphthalene-1-sulfonamide

2934.99.9001

Powder      -20°C    3 years

                     4°C     2 years

In solvent   -80°C    6 months

                  -20°C    1 month

Note: (1). This product requires protection from light (avoid light exposure) during transportation and storage.  (2). Please store this product in a sealed and protected environment (e.g. under nitrogen), avoid exposure to moisture.

Room temperature (This product is stable at ambient temperature for a few days during ordinary shipping and time spent in Customs)

DMSO : ~62.5 mg/mL (~186.31 mM)
H2O : ~1 mg/mL (~2.98 mM)

Note: Listed below are some common formulations that may be used to formulate products with low water solubility (e.g. < 1 mg/mL), you may test these formulations using a minute amount of products to avoid loss of samples.

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Injection Formulation 1: DMSO : Tween 80： Saline = 10 : 5 : 85 (i.e. 100 μL DMSO stock solution → 50 μL Tween 80 → 850 μL Saline)
*Preparation of saline: Dissolve 0.9 g of sodium chloride in 100 mL ddH ₂ O to obtain a clear solution.
Injection Formulation 2: DMSO : PEG300 ：Tween 80 : Saline = 10 : 40 : 5 : 45 (i.e. 100 μL DMSO → 400 μLPEG300 → 50 μL Tween 80 → 450 μL Saline)
Injection Formulation 3: DMSO : Corn oil = 10 : 90 (i.e. 100 μL DMSO → 900 μL Corn oil)
Example: Take the Injection Formulation 3 (DMSO : Corn oil = 10 : 90) as an example, if 1 mL of 2.5 mg/mL working solution is to be prepared, you can take 100 μL 25 mg/mL DMSO stock solution and add to 900 μL corn oil, mix well to obtain a clear or suspension solution (2.5 mg/mL, ready for use in animals).

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Injection Formulation 4: DMSO : 20% SBE-β-CD in saline = 10 : 90 [i.e. 100 μL DMSO → 900 μL (20% SBE-β-CD in saline)]
*Preparation of 20% SBE-β-CD in Saline (4°C，1 week): Dissolve 2 g SBE-β-CD in 10 mL saline to obtain a clear solution.
Injection Formulation 5: 2-Hydroxypropyl-β-cyclodextrin : Saline = 50 : 50 (i.e. 500 μL 2-Hydroxypropyl-β-cyclodextrin → 500 μL Saline)
Injection Formulation 6: DMSO : PEG300 : castor oil : Saline = 5 : 10 : 20 : 65 (i.e. 50 μL DMSO → 100 μLPEG300 → 200 μL castor oil → 650 μL Saline)
Injection Formulation 7: Ethanol : Cremophor : Saline = 10： 10 : 80 (i.e. 100 μL Ethanol → 100 μL Cremophor → 800 μL Saline)
Injection Formulation 8: Dissolve in Cremophor/Ethanol (50 : 50), then diluted by Saline
Injection Formulation 9: EtOH : Corn oil = 10 : 90 (i.e. 100 μL EtOH → 900 μL Corn oil)
Injection Formulation 10: EtOH : PEG300：Tween 80 : Saline = 10 : 40 : 5 : 45 (i.e. 100 μL EtOH → 400 μLPEG300 → 50 μL Tween 80 → 450 μL Saline)

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Oral Formulation 1: Suspend in 0.5% CMC Na (carboxymethylcellulose sodium)
Oral Formulation 2: Suspend in 0.5% Carboxymethyl cellulose
Example: Take the Oral Formulation 1 (Suspend in 0.5% CMC Na) as an example, if 100 mL of 2.5 mg/mL working solution is to be prepared, you can first prepare 0.5% CMC Na solution by measuring 0.5 g CMC Na and dissolve it in 100 mL ddH2O to obtain a clear solution; then add 250 mg of the product to 100 mL 0.5% CMC Na solution, to make the suspension solution (2.5 mg/mL, ready for use in animals).

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Oral Formulation 3: Dissolved in PEG400
Oral Formulation 4: Suspend in 0.2% Carboxymethyl cellulose
Oral Formulation 5: Dissolve in 0.25% Tween 80 and 0.5% Carboxymethyl cellulose
Oral Formulation 6: Mixing with food powders

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Note: Please be aware that the above formulations are for reference only. InvivoChem strongly recommends customers to read literature methods/protocols carefully before determining which formulation you should use for in vivo studies, as different compounds have different solubility properties and have to be formulated differently.

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 (Please use freshly prepared in vivo formulations for optimal results.)