Doxycycline calcium

Cat. No.: HY-N0565C: Data Sheet Handling Instructions
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Doxycycline calcium is an orally active highly lipophilic, tissue-permeable MMP inhibitor with broad-spectrum antibacterial activity. Doxycycline calcium is also a semi-synthetic antibiotic with chelating properties, which blocks bacterial protein synthesis and inhibits extracellular matrix degradation through interactions with zinc and calcium atoms. Doxycycline calcium also inhibits mitochondrial biogenesis, translation, and the expression of respiratory chain proteins. Doxycycline calcium induces apoptosis, inhibits autophagy and EMT, downregulates stem cell markers, and activates the PI3K-AKT pathway, thereby effectively inhibiting the viability and proliferation of cancer cells such as breast cancer cells. Doxycycline calcium also promotes the survival and self-renewal of embryonic stem cells and neural stem cells, and reduces the frequency of medium changes in culture. Doxycycline calcium has been applied in studies related to breast cancer, prostate cancer, bladder cancer, and other cancers.

For research use only. We do not sell to patients.

CAS No. : 94088-85-4

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Other In-stock Forms of Doxycycline calcium:

Doxycycline Doxycycline hydrochloride Doxycycline hyclate Doxycycline monohydrate

Other Forms of Doxycycline calcium:

Doxycycline-d₃ (hyclate) (major) In-stock
Doxycycline In-stock
Doxycycline hydrochloride In-stock
Doxycycline hyclate In-stock
Doxycycline monohydrate In-stock

173 Publications Citing Use of MCE Doxycycline calcium

In Vivo Efficacy Study

IHC

In Vivo Imaging

Cell Imaging/Staining

RT-PCR

ELISA

Microbiological Assay

: Doxycycline calcium purchased from MedChemExpress. Usage Cited in: Cancer Commun (Lond). 2025 May;45(5):608-631. [Abstract]; Bioluminescence imaging of the lung metastasis burden after inoculation of MKN45 cells with DOX (500 mg/kg, daily, 30 days)-induced CagA expression and MA treatment.

: Doxycycline calcium purchased from MedChemExpress. Usage Cited in: Cancer Commun (Lond). 2025 May;45(5):608-631. [Abstract]; After inoculating MKN45 cells with DOX-induced CagA expression and MA treatment, HE staining was performed on lung tissue sections of a specified group. The quantitative results of the number of lung metastases are shown on the right.

: Doxycycline calcium purchased from MedChemExpress. Usage Cited in: Nat Cell Biol. 2025 Apr;27(4):641-653. [Abstract]; HeLa cells stably expressing tet-on-shScramble (Scr) or tet-on-shMAT2A were pretreated with Doxycycline hydrochloride (Dox, 1 μg/ mL) for two days, and the cell lysates were analysed by immunoblotting with the indicated antibodies.

: Doxycycline calcium purchased from MedChemExpress. Usage Cited in: Chem Eng J. 2025 Oct 8;524:169421.; Therapeutic effect of VRC, Doxycycline hydrochloride (Doxy, 0.05 %, 5 μl, 4 times per day for 5 days) on DXMS- associated severe FK. Dynamic phenotype of corneas with different groups observed by slit lamp on each day after infection (n = 6/group).

: Doxycycline calcium purchased from MedChemExpress. Usage Cited in: J Exp Clin Cancer Res. 2025 Mar 19;44(1):99. [Abstract]; Nude mice (nu/nu) were surgically engrafted intracranially with GSCs labeled with luciferase and transduced with a Doxycycline hydrochloride (DOX)-inducible system for shPHGDH expression. Mice harboring either GSC19 (A) or GSC29 (B) were randomly assigned to treatment groups (6 mice per group). Starting from day 7 or day 9 post-engraftment, as per the study design (A, B, left), treatments were administered: control, 3 Gy ionizing radiation (IR) delivered weekly over three sessions, 2 mg/mL DOX in the drinking water or a combination of IR and DOX. The progression of GBM xenografts was monitored through bioluminescence imaging, and representative images are displayed (A, B, right).

: Doxycycline calcium purchased from MedChemExpress. Usage Cited in: Cell Rep. 2025 Nov 25;44(11):116496. [Abstract]; MOLM-13 cells were treated with Doxycycline hydrochloride (25 µM) or Chloramphenicol (25 µM) for 24 h. Mitochondrial translation and MMP were examined (n=3)

: Doxycycline calcium purchased from MedChemExpress. Usage Cited in: AMB Express. 2024 Dec 24;14(1):141. [Abstract]; Combinational antimicrobial effects between PVB and conventional antibiotics against MRSA ATCC 43,300. TET, Tetracycline. DOX, Doxycycline. E, Erythromycin. AZI, Azithromycin. P, Penicillin. CAZ, Ceftazidime. AMP, Ampicillin. CEZ, Cefazolin. CEF, Cefotaxime. OXA, Oxacillin. CRO, Ceftriaxone. AMK, Amikacin. GEN, Gentamycin, KANA, Kanamycin. TOB, Tobramycin. SPC, Spectinomycin.

: Doxycycline calcium purchased from MedChemExpress. Usage Cited in: Nat Genet. 2024 Feb;56(2):294-305. [Abstract]; RT-qPCR analysis of STAT5A and target gene expression in STB-BL with DOX (5 μM, 6 days)-inducible overexpression of STAT5A.

: Doxycycline calcium purchased from MedChemExpress. Usage Cited in: Nat Genet. 2024 Feb;56(2):294-305. [Abstract]; RT-qPCR analysis of MITF and target gene expression in STB-BL with DOX (5 μM, 6 days)-inducible overexpression of MITF.

: Doxycycline calcium purchased from MedChemExpress. Usage Cited in: Nat Genet. 2024 Feb;56(2):294-305. [Abstract]; ELISA analysis of CSH2 expression in trophoblast organoids derived from hTSCs-BL with DOX (5 μM, 6 days)-inducible overexpression of MITF.

: Doxycycline calcium purchased from MedChemExpress. Usage Cited in: Nat Genet. 2024 Feb;56(2):294-305. [Abstract]; Bright fields and immunofluorescence analysis of trophoblast markers (CDH1 and hCG) in trophoblast organoids derived from hTSCs-BL with DOX (5 μM, 6 days)-inducible overexpression of MITF.

: Doxycycline calcium purchased from MedChemExpress. Usage Cited in: Cancer Commun (Lond). 2024 Feb;44(2):251-272. [Abstract]; The expression of NHE7 in Dox-inducible NHE7-mAID-KO MHCC97L cells with or without doxycycline (DOX, 2 mg/mL) and indole-3-acetic acid (IAA, 50 μg/mL) treatment for 24 h was examined by immunoblotting.

: Doxycycline calcium purchased from MedChemExpress. Usage Cited in: Cancer Commun (Lond). 2024 Feb;44(2):251-272. [Abstract]; Mice orthotopically injected with NHE7-mAID-KO MHCC97L cells were administered vehicle (DMSO), DOX (2 mg/mL in drinking water containing 5% sucrose), IAA (25 mg/kg/day) and sorafenib (30 mg/kg/day) for 2 weeks (n = 6). Bioluminescence imaging of whole mice was performed at the end of the experiment, and the signal intensity was quantified.

: Doxycycline calcium purchased from MedChemExpress. Usage Cited in: Cancer Commun (Lond). 2024 Feb;44(2):251-272. [Abstract]; Mice orthotopically injected with NHE7-mAID-KO MHCC97L cells were administered vehicle (DMSO), DOX (2 mg/mL in drinking water containing 5% sucrose), IAA (25 mg/kg/day) and sorafenib (30 mg/kg/day) for 2 weeks (n = 6). Immunohistochemistry of NHE7 and Rab21 in liver tumor tissues.

: Doxycycline calcium purchased from MedChemExpress. Usage Cited in: Mol Cancer. 2020 Sep 9;19(1):139. [Abstract]; ATL cells MT1 stably transfected with a TET-inducible FBXW7 wild type or W425R were induced with doxycycline (2 μg/ml, 24 h) and degradation (WT) or lack thereof (W425R) of endogenous BRAF was confirmed by Western blot.

: Doxycycline calcium purchased from MedChemExpress. Usage Cited in: Mol Cancer. 2020 Sep 9;19(1):139. [Abstract]; ATL cells MT1 stably transfected with a TET-inducible FBXW7 wild type, endogenous degradation of BRAF by wild type FBXW7 was confirmed by immunohistochemistry following induction of FBXW7 with doxycycline (2 μg/ml, 24 h).

: Doxycycline calcium purchased from MedChemExpress. Usage Cited in: Cancer Lett. 2016 Jun 28;376(1):188-96. [Abstract]; Doxycycline inducible shRNA can effectivly knockdown the expression of AKT in HCC cells. Cells are treated with 100ng/mL Doxcycline and cultured for the indicated times.

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Biological Activity
Purity & Documentation
References
Customer Review

Description

IC₅₀ & Target

Tetracycline

In Vitro

Doxycycline (11.39 μM + MCF-7; 7.13 μM + MDA-MB-468; 72 h) calcium significantly reduces the population of CD44⁺/CD24^-/low breast cancer stem cells in MCF-7 and MDA-MB-468 cells^[2].
Doxycycline (11.39 μM+MCF-7; 7.13 μM+MDA-MB-468; 72 h) calcium significantly downregulates the mRNA and protein expression of stem cell markers (Nanog, Oct4, Sox2, c-Myc, CD44) in MCF-7 and MDA-MB-468 breast cancer cells^[2].
Doxycycline (1 μg/mL; 5 d) calcium increases the colony formation efficiency of undifferentiated human embryonic stem cells (H9, HSF6, H1, H7, HUES6) and human induced pluripotent stem cells (retrovirus-derived, lentivirus-derived, protein-induced lines), and elevates the number of colonies expressing undifferentiated markers under various culture conditions^[3].
Doxycycline (1 μg/mL; 5 d) calcium promotes the survival and proliferation of H9 human embryonic stem cells (hESC) more effectively than Y-27632 (HY-10071), resulting in larger cell colonies, a higher proportion of undifferentiated AP⁺ cells, as well as downregulated expression of pro-apoptotic genes and upregulated expression of pluripotency-related genes^[3].
Doxycycline (1 μg/mL; 6 d; passage 12) calcium reduces cell apoptosis, increases S-phase cell accumulation, supports the long-term expansion of H9 human embryonic stem cells (H9 hESCs) and Lenti-1 induced pluripotent stem cells (Lenti-1 hiPSCs) cultured in clusters on MEF feeder layers, and maintains the pluripotency and normal karyotype of the cells during repeated passaging^[3].
Doxycycline (1 μg/mL; 7 d) calcium maintains the viability, proliferation and expression of undifferentiated markers of H9 human embryonic stem cells for up to 7 days without medium change, and reduces apoptotic cell death of newly seeded dissociated cells and preformed cell clusters within 3 days without medium change^[5].
Doxycycline (1 μg/mL; 30-72 d) calcium supports long-term subculture of H9, HSF6 human embryonic stem cells as well as Retro-1, Lenti-1, Pro-1 human induced pluripotent stem cells. The culture medium is replaced every 3 days, and the growth rate remains consistent with that under the standard daily medium change condition during passages 5 to 12^[5].
Doxycycline (1 μg/mL; 72 d) calcium maintains normal karyotype, expression of undifferentiated markers and pluripotency (the ability to differentiate into all cells of the three germ layers in vitro) of H9 human embryonic stem cells that are subcultured 12 times with medium replacement every 3 days^[5].

MedChemExpress (MCE) has not independently confirmed the accuracy of these methods. They are for reference only.

Cell Viability Assay^[2]

Cell Line:	LNT-229, G55, and U343 glioma cells
Concentration:	0.01, 0.1, 1 or 10 µg/mL
Incubation Time:	4 days
Result:	Affected growth of glioma cells only under high concentration (10 µg/mL).

Cell Viability Assay^[2]

Cell Line:	SVG cells
Concentration:	0.01, 0.1, 1 or 10 µg/mL
Incubation Time:	24 hours
Result:	Decreaseed MT-CO1 protein content with concentrations of 1 µg/mL and higher.

Cell Proliferation Assay^[4]

Cell Line:	MCF 12A, 293T cells
Concentration:	100 ng/mL, 1 µg/mL
Incubation Time:	96 hours
Result:	Caused reduced proliferation of MCF 12A and 293T cells at 1 µg/mL.

Cell Viability Assay^[5]

Cell Line:	MCF-7, MDA-MB-468 cells
Concentration:	0-250 μM
Incubation Time:	72 hours
Result:	Inhibited breast cancer cells in a dose-dependent manner with IC₅₀ values for MCF-7 and MDA-MB-468 of 11.39 μM and 7.13 μM respectively.

In Vivo

Doxycycline (25-50 mg/kg; p.o.; twice daily; 10 days) calcium exerts dose-dependent toxic effects on male Wistar rats: a dosage of 50 mg/kg twice daily for 10 consecutive days causes 30% mortality, cardiomyopathy, and significant elevation of biomarkers for muscle and cardiac injury; whereas a dosage of 25 mg/kg twice daily for 10 consecutive days only induces mild skeletal muscle injury without cardiac toxicity^[1].
Doxycycline (60 mg/kg; intraperitoneal injection; once daily for 15 consecutive days) calcium significantly inhibits the growth of CD44v9-expressing prostate cancer xenografts by reducing the proliferation of CD44v9-positive cells^[4].
Supplementary culture of H9 human embryonic stem cells (H9 hESCs) with Doxycycline (1 μg/mL; medium changed every 3 days; up to passage 12) calcium maintains their pluripotency and enables in vivo teratoma formation, which contains all three embryonic germ layers^[5].

Note:
Please do not refer to only one article to determine the experimental conditions. It is recommended to determine the optimal experimental conditions (animal strain, age, dosage, frequency and cycle, detection time and indicators, etc.) through preliminary experiments before the formal experiment.

Doxycycline (oral gavage; 200 or 800 mg/kg; once daily; 3 months) reduces MMP-9 activity in untreated HT mice in a dose-dependent manner^[3].
Doxycycline and Tetracycline (HY-A0107), act systemically after absorption from the upper gastrointestinal tract. The main advantage of Doxycycline over Tetracycline is its longer activity, and it can be taken twice or once a day. The peak concentration of both agents is similar, but in the case of Doxycycline the time to peak concentration is shorter, and half life is significantly longer^[6].

Doxycycline (Dox) is often used as an inducer in molecular biology studies to induce gene expression. In cells or model animals that have constructed tetracycline induced expression systems (Tet-On/Tet-Off systems), the expression of target genes can be precisely controlled by adding or removing Dox^[7]^[8].
Dose reference for Dox induction^[7]^[8]:
(1) Model animal: male Sprague-Dawley rats
Tet regulatory system: 20-3000 ppm of Dox is supplied in diet.
(2) Model animal: Cags mice
Tet regulatory system: 625 ppm of Dox is supplied in diet.

Induction of Modeling ON-OFF System (Gene expression regulation)^[6]^[7]^[8]

Background

Doxycycline is often used as an inducer in molecular biology research to induce gene expression. In cells or model animals that have constructed a Tetracycline (Tet; HY-A0107) inducible expression (Tet-ON/Tet-OFF) system, the expression of the target gene can be precisely controlled by adding or removing Doxycycline. Doxycycline can act as an inhibitor of transcriptional activation in the Tetracycline (Tc)-controlled transactivation (tTA) system, and as an inducer of transcriptional activation in the "reverse tTA' system. Doxycycline and Tetracycline both act systemically after being absorbed by the upper gastrointestinal tract. In comparison, the main advantage of Doxycycline is that it has a longer activity and can be taken twice or once a day. Although the peak concentrations of the two are similar, Doxycycline takes a shorter time to reach peak concentration and has a significantly longer half-life.

Specific Modeling Methods

Rat^[8]: Sprague-Dawley rats • male • adult middle-aged (12-month-old)
Administration: (for GDNF as targeted gene) 3g/kg (dietary with regular food) • po • once daily for 6 days

Note

(1) Recommend use the recombinant adeno-associated virus (rAAV)-based bicistronic tetracycline (tet)-OFF construct was used for dynamic control of GDNF (target gene) expression during long-term expression^[7].
(2) 3 g/kg dietary DOX produced DOX serum levels equivalent to 1mg/ml DOX in drinking water.

Modeling Indicators

Molecular changes: The expression level of the target gene decreases.

Phenotype changes: The positively correlated phenotype corresponding to the target gene is alleviated.

MedChemExpress (MCE) has not independently confirmed the accuracy of these methods. They are for reference only.

Animal Model:	6-month-old female Heterozygous Col3a1-deficient (HT) mice^[3]
Dosage:	200 or 800 mg/kg
Administration:	Oral gavage; 200 or 800 mg/kg; once daily; 3 months
Result:	Reduced MMP-9 activity in a dose-dependent manner.

Molecular Weight

524.59

Formula

C₂₂H₂₄Ca₂N₂O₈

CAS No.

94088-85-4

SMILES

O=C(C(C1=O)=C(O)[C@@H](N(C)C)[C@]2([H])[C@@H](O)[C@]3([H])[C@@H](C)C4=C(C(C3=C(O)[C@@]21O)=O)C(O)=CC=C4)N.[Ca].[Ca]

Shipping

Room temperature in continental US; may vary elsewhere.

Storage

Please store the product under the recommended conditions in the Certificate of Analysis.

Purity & Documentation

Handling Instructions (2659 KB)

References

[1]. El-Neweshy MS, et al. Experimental doxycycline overdose in rats causes cardiomyopathy. Int J Exp Pathol. 2013;94(2):109-114. [Content Brief]

[2]. Zhang L, et al. Doxycycline inhibits the cancer stem cell phenotype and epithelial-to-mesenchymal transition in breast cancer. Cell Cycle. 2017;16(8):737-745. [Content Brief]

[3]. Chang M Y, Rhee Y H, Yi S H, et al. Doxycycline enhances survival and self-renewal of human pluripotent stem cells[J]. Stem cell reports, 2014, 3(2): 353-364.

[4]. Matsumoto T, et al. Doxycycline induces apoptosis via ER stress selectively to cells with a cancer stem cell-like properties: importance of stem cell plasticity[J]. Oncogenesis, 2017, 6(11): 397.

[5]. Chang MY, et al. Doxycycline supplementation allows for the culture of human ESCs/iPSCs with media changes at 3-day intervals. Stem Cell Res. 2015;15(3):608-613. [Content Brief]

[6]. Manfredsson FP, et al. Tight Long-term dynamic doxycycline responsive nigrostriatal GDNF using a single rAAV vector. Mol Ther. 2009 Nov;17(11):1857-67. [Content Brief] [Content Brief]

[7]. Kistner A, et al. Doxycycline-mediated quantitative and tissue-specific control of gene expression in transgenic mice. Proc Natl Acad Sci U S A. 1996 Oct 1;93(20):10933-8. [Content Brief]

[8]. Niv Y. Doxycycline in Eradication Therapy of Helicobacter pylori--a Systematic Review and Meta-Analysis. Digestion. 2016;93(2):167-73. [Content Brief]