Doxycycline (hyclate) [24390-14-5]

Commande minimum 2

Référence NB-64-07882-1g

Conditionnement : 1g

Marque : Neo Biotech


Doxycycline (hyclate) (Synonyms: WC2031, Doxycycline hydrochloride hemiethanolate hemihydrate, Doxycycline hyc...)

Catalog No. T1687L Copy Product Info
Purity: 99.94%
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Doxycycline (hyclate) is an orally active tetracycline antibiotic with broad-spectrum inhibitory activity against matrix metalloproteinases (MMPs), possessing both antibacterial and antitumor properties, and is commonly used in inducible gene expression ON-OFF systems.

Doxycycline (hyclate)

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Synonyms WC2031, Doxycycline hydrochloride hemiethanolate hemihydrate, Doxycycline hyclate

Doxycycline (hyclate) is an orally active tetracycline antibiotic with broad-spectrum inhibitory activity against matrix metalloproteinases (MMPs), possessing both antibacterial and antitumor properties, and is commonly used in inducible gene expression ON-OFF systems.

Doxycycline (hyclate)
Cas No. 24390-14-5
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Purity:99.94%
Color:Yellow
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Product Introduction

Bioactivity
Description
Doxycycline (hyclate) is an orally active tetracycline antibiotic with broad-spectrum inhibitory activity against matrix metalloproteinases (MMPs), possessing both antibacterial and antitumor properties, and is commonly used in inducible gene expression ON-OFF systems.
Targets&IC50
NCI-H446 cells:1.70 μM, Pancreatic Cancer cells (48 h):987.5 μM, Pancreatic Cancer cells (72 h):99.64 μM, A549 cells:1.06 μM, chlamydial isolates:0.497 μg/mL, Pancreatic Cancer cells (96 h):50.02 μM
In vitro
METHODS: Twenty-two tumor cells were treated with Doxycycline for 48 h. Cell viability was measured by MTT.
RESULTS: Lung cancer cells were more sensitive to Doxycycline than most other cell lines. NCI-H446 and A549 cells were sensitive to Doxycycline with IC50s of 1.70 μM and 1.06 μM, respectively. [1]
METHODS: Human pancreatic cancer cells PANC-1 were treated with Doxycycline (10-40 μg/mL) for 8-24 h. The cell cycle was detected by Flow Cytometry.
RESULTS: Doxycycline caused the cells to arrest in the G1-S phase of the cell cycle. [2]
In vivo
METHODS: To detect anti-tumor activity in vivo, Doxycycline (50 mg/kg) was administered intraperitoneally once daily for fifteen days to BALB/c mice harboring human mammary carcinoma tumor 4T1.
RESULTS: Doxycycline delayed tumor growth and reduced the number of tumor-associated macrophages and blood vessels. [3]
METHODS: To assay antimicrobial activity in vivo, Doxycycline (40 mg/kg) was administered intraperitoneally to BALB/c mice exposed to B. pseudomallei 1026b twice daily for two weeks.
RESULTS: Doxycycline-treated mice survived 100% of the time, and no B. pseudomallei DNA was amplified from the lungs or spleens of the majority of the surviving mice. [4]
Disease Modeling Protocol
Lung airway-specific genetically engineered disease model
  • Modeling Mechanism:

    Doxycycline (hyclate) binds to rtTA via the Tet-On system, activating transgene expression downstream of the TRE promoter in airway epithelial cells. Aerosol delivery allows for rapid, controllable, dose-dependent gene induction locally in the lung airways, enabling the simulation or study of functional changes in lung disease-related genes within specific time windows.

  • Related Products:

    Doxycycline (hyclate) (T1687L)

  • Modeling Method:

    Experimental Subject:

    Mice, Transgenic mice (Rosa26-rtTA, CK5-rtTA, tet(O)H2BGFP, Tet(O)Cre, R26-LSL-NICD), Background strains: C57/BL6, FVB/N

    Dosage and Administration Route:

    Mice anaesthetised and placed in 50 ml centrifuge tubes with nasal openings, then placed in micro-isolation cages for inhalation of nebulised 50 mg/mL Doxycycline; control: nebulised PBS

    Dosing Frequency and Duration Model:

    Three cycles (20 minutes inhalation+20-minute interval)

  • Validation:

    Immunofluorescence, β-gal, or fluorescent reporter gene assays showed that the transgene was specifically expressed in the lung airway epithelium; qPCR and Western blot confirmed that transgene expression was rapidly upregulated after Doxycycline administration; transgene expression gradually decreased after Doxycycline withdrawal, indicating that the model has good reversibility and time controllability.

*Precautions: In all experiments, mice were killed within 24 hours after final inhalation for transgene expression analysis.

*References:Tata PR,et,al. Airway-specific inducible transgene expression using aerosolized doxycycline. Am J Respir Cell Mol Biol. 2013 Dec;49(6):1048-56.

Kinase Assay
Gelatin (0.1% (w/v) is added to standard LaemmLi acrylamide polymerization mixture. Tissue extract is mixed 1:2 with sample buffer [250 mM Tris-Cl pH 6.8, 10% (w/v) SDS, 20% (v/v) glycerol, 0.005% (w/v) bromphenol blue]. Serum is diluted 1:10 with electrophoresis buffer (2.5 mM Tris, 20 mM glycine, 0.005% SDS) and mixed 1:2 with sample buffer. Twenty μLs are loaded after 10-min incubation at room temperature without boiling. After electrophoresis at 90 V, the gels are soaked in 2.5% (w/v) Triton X-100, incubated 2 to 3 days at 37°C in gelatin digestion buffer [50 mM Tris-Cl, pH 8.0, 8 mM CaCl2, 10 mM ZnSO2, 0.02% (w/v) NaN3], stained in 0.05% Coomassie blue R-250 in acetic acid/methanol/water (1:4.5:4.5 by volume), destained in 10% acetic acid and 5% methanol, and scanned for lysis band intensity. The lysis band intensity is proportional to gelatinase activity and is quantified densitometrically by using One-Dimensional Scan software. The result, a number between 0.07 and 3.75, is normalized to the protein content by dividing the densitometry result with the relative optical density from the BCA protein assay kit result. The result is used for the analysis as the arbitrary unit. For the total MMP activity results of lysis bands of pro-MMP-9, active MMP-9, pro-MMP-2, and active MMP-2 are added. A protein size marker is used to determine the correct size.
Cell Research
Doxycycline is diluted in culture medium at a concentration of 10 μg/mL. All in vitro treatments are performed in SMC cultures at 90% confluence, when ECM synthesis in SMC starts to be evident. Doxycycline (20 nM) is diluted in culture medium at a concentration of 10 μg/mL (20 nM), at which no toxicity or variation in primary cultured SMC proliferation has been reported, as well as in other cell lines and the incubation time is 48 h. SMC-C and SMC-Ch are seeded at equal cell density in 6-well plates and, when confluence reaches 90%, 1 mL culture medium is added to each well containing 3.7×104?Bq L-[5-3H]-proline (9.62×1011 Bq/mmol). After 48?h incubation, cells are lysed with 0.5 mL 0.5 mol/L NaOH for 1 h. The resulting solution is neutralized with an equal amount of 0.5 mol/L HCl, and 50 μL are used to measure total proteins with the Bradford method. One volume of 10% TCA is added to the remaining 250 μL and centrifuged at 13,000 g for 15?min at 4°C. The resulting precipitate is dissolved in 100 μL 0.2 mol/L NaOH, and then neutralized with 1 mol/L HCl. The solution is incubated with collagenase buffer (Tris-HCl, pH 7.6 20 mM, and CaCl2 250 mM final concentration) and 10 units of collagenase at 37°C overnight. Then, 150 μL 10% TCA are added and centrifuged at 13000 g for 15?min at 4°C. The resulting supernatant is added to 4 mL of scintillation fluid and the radioactivity is measured in a liquid scintillation counter LS 600 TA.
SynonymsWC2031, Doxycycline hydrochloride hemiethanolate hemihydrate, Doxycycline hyclate
Chemical Properties
Molecular Weight512.90
FormulaC22H24N2O8·HCl·1/2C2H6O·1/2H2O
Cas No.24390-14-5
SmilesO.Cl.Cl.CCO.C[C@@H]1[C@H]2[C@H](O)[C@H]3[C@H](N(C)C)C(=O)C(C(N)=O)=C(O)[C@@]3(O)C(=O)C2=C(O)c2c(O)cccc12.C[C@@H]1[C@H]2[C@H](O)[C@H]3[C@H](N(C)C)C(=O)C(C(N)=O)=C(O)[C@@]3(O)C(=O)C2=C(O)c2c(O)cccc12
Relative Density.no data available
Storage & Solubility Information
StoragePowder: -20°C for 3 years | In solvent: -80°C for 1 year Shipping with blue ice/Shipping at ambient temperature.
Solubility Information
H2O: 13.89 mg/mL (27.08 mM), Sonication is recommended.
DMSO: 250 mg/mL (487.42 mM), Sonication is recommended.
Solution Preparation Table
H2O/DMSO
1mg5mg10mg50mg
1 mM1.9497 mL9.7485 mL19.4970 mL97.4849 mL
5 mM0.3899 mL1.9497 mL3.8994 mL19.4970 mL
10 mM0.1950 mL0.9748 mL1.9497 mL9.7485 mL
20 mM0.0975 mL0.4874 mL0.9748 mL4.8742 mL
DMSO
1mg5mg10mg50mg
50 mM0.0390 mL0.1950 mL0.3899 mL1.9497 mL
100 mM0.0195 mL0.0975 mL0.1950 mL0.9748 mL
Note : The dilution table applies only to solid products. For liquid products, please calculate the stock solution based on the stated concentration and/or density.

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