Doxycycline hyclate dox

Manufactured by Merck Group

Sourced in United States

Doxycycline hyclate (Dox) is a broad-spectrum antibiotic used in various laboratory applications. It functions as an inhibitor of bacterial protein synthesis.

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Lab products found in correlation

Alexa fluor 488, by Thermo Fisher Scientific (3 mentions) Mouse anti ha 11 clone 16b12, by BioLegend (3 mentions) Mouseanti ha 11 clone 16b12, by Thermo Fisher Scientific (3 mentions) Dulbecco modified eagle medium (dmem), by Thermo Fisher Scientific (3 mentions) Phalloidin af647, by Thermo Fisher Scientific (2 mentions) Hybridomacells secreting mouse monoclonal anti iav np clone h16 l10 4r5, by American Type Culture Collection (2 mentions) Polyclonal rabbit anti ha y 11, by Santa Cruz Biotechnology (2 mentions) Fetal bovine serum (fbs), by Thermo Fisher Scientific (2 mentions) Pen strep, by Thermo Fisher Scientific (2 mentions) N acetylcysteine nac, by Merck Group (2 mentions) Bafilomycin a1, by Merck Group (2 mentions) 17β estradiol e2, by Merck Group (1 mentions) P iκbα, by Cell Signaling Technology (1 mentions) P p65 s536, by Cell Signaling Technology (1 mentions) Ck8 18, by Cell Signaling Technology (1 mentions) E cadherin, by Cell Signaling Technology (1 mentions) Ab53115, by Abcam (1 mentions) Tnf α, by R&D Systems (1 mentions) Vimentin, by Cell Signaling Technology (1 mentions) Mdv3100, by Santa Cruz Biotechnology (1 mentions)

Spelling variants of this product

Doxycycline (1805 citations) Doxycycline hyclate (325 citations) Doxycycline (DOX) (189 citations) DOX hyclate (17 citations) Doxycycline (doxycycline hyclate, (3 citations)

37 protocols using doxycycline hyclate dox

Monoclonal Anti-IAV NP Antibody Purification

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Doxycycline hyclate (Dox) was purchased from Sigma Aldrich. Hybridoma
cells secreting mouse monoclonal anti-IAV NP (clone H16-L10-4R5, ATCC
HB-65)^{25 (link)} were obtained
from ATCC and antibodies in the supernatant purified using a protein G column.
Polyclonal rabbit anti-neomycin phosphotransferase II (NPTII) was purchased from
Fitzgerald Industries International. Mouse anti-HA.11 (clone 16B12) was acquired
from BioLegend, polyclonal rabbit anti-HA (Y-11) from Santa Cruz. Mouse
anti-HA.11 (clone 16B12) coupled to Alexa Fluor (AF) 488 or AF594, as well as
fluorescently-labeled secondary antibodies and AF647 Phalloidin were obtained
from Life Technologies.

Schmidt F.I., Hanke L., Morin B., Brewer R., Brusic V., Whelan S.P, & Ploegh H.L. (2016). Phenotypic lentivirus screens to identify functional single domain antibodies. Nature microbiology, 1(8), 16080.

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Regulation of Epithelial-Mesenchymal Transition

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17β-estradiol (E2) and doxycycline hyclate (DOX) were purchased from Sigma. TNFα was purchased from R&D Systems. ICI and IKK7 were purchased from Sigma (#I4409) and SelleckChem (#S2882), respectively. Antibodies for IKKβ (#2370), IKKα (#2682), p-IκBα (#2859), IκBα (#4814), p-p65 S536 (#3033), ERα (#8644), E-cadherin (#3195P), vimentin (#5741P), CK8/18 (#4546), GATA3 (#5852) and P63-α (#4892) were purchased from Cell Signaling. Antibodies for p65 (#sc-372) and Fibronectin (#sc-9068) were purchased from Santa Cruz. Antibodies for FoxA1 (#ab55178), and CK14 (#ab53115) were purchased from Abcam. Antibodies for CK5 (#CK5-L-CE) and β-actin (#A5441) were purchased from, Leica and Sigma, respectively.

El-Shennawy L., Dubrovskyi O., Kastrati I., Danes J.M., Zhang Y., Whiteley H.E., Creighton C.J, & Frasor J. (2017). Coactivation of estrogen receptor and IKK-β induces a dormant metastatic phenotype in ER-positive breast cancer. Cancer research, 78(4), 974-984.

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Optimizing Compound Delivery for Cell Studies

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DHT (4,5α-Dihydrotestosterone) was purchased from Sigma-Aldrich (St. Louis, MO, USA; A8380) and dissolved in ethanol at 10 μM to be used at a final concentration of 10 nM. MDV3100 (enzalutamide), from Santa Cruz Biotechnology (Dallas, TX, USA; sc-364354), was diluted in DMSO at 10 mM and used at 10µM. GSK126 (EZH2 inhibitor) was purchased from MedChemExpress (Monmouth Junction, NJ, USA; HY-13470), dissolved in DMSO, and used in a range of concentrations for 24 h. C2 Ceramide (N-acetoyl-D-erythro-sphingosine) was purchased from Avanti Polar Lipids (Alabaster, AL, US; 860502), dissolved in DMSO, and used at a final concentration of 20 μM. Doxycycline Hyclate (Dox, used at a final concentration of 0.25 μg/mL) and C16 Ceramide 1-phosphate (C1P, 860533P) were purchased from Sigma Aldrich (St. Louis, MO, US). An aqueous dispersion (in the form of liposomes) was prepared by sonicating C1P (1 mg) in sterile nanopure water (600 μL) on ice using a probe sonicator for six cycles of 8 s on and 5 s off until a clear dispersion was obtained. C1P concentration in the stock solution was 2.6 mM and a final concentration of 20 μM was used for all cellular assays. This procedure is considered preferable to C1P dispersions in organic solvents because lipid droplet formation is minimized and exposure of cells to alcohols or dodecane is avoided.

Camacho L., Zabala-Letona A., Cortazar A.R., Astobiza I., Dominguez-Herrera A., Ercilla A., Crespo J., Viera C., Fernández-Ruiz S., Martinez-Gonzalez A., Torrano V., Martín-Martín N., Gomez-Muñoz A, & Carracedo A. (2021). Identification of Androgen Receptor Metabolic Correlome Reveals the Repression of Ceramide Kinase by Androgens. Cancers, 13(17), 4307.

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Doxycycline-Loaded Halloysite Nanotubes

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Doxycycline hyclate (DOX, Sigma-Aldrich, St. Louis, MO, USA) was dissolved in 5 mL of 50 mM HEPES buffer (pH = 7.2, Fisher Scientific, Pittsburgh, PA, USA) at three distinct concentrations (w/v): 10% (0.5 g), 20% (1 g), and 30% (1.5 g) by stirring (Fisher Thermix, model 310T, Fisher Scientific). The loading process started by mixing a predetermined amount (1.25 g) of pre-sieved (<45 µm) Halloysite^® nanotubes (HNT, Dragonite 1415JM, Applied Minerals Inc., New York, NY, USA) in 5 mL of each DOX solution and followed by sonication for 2 h [19 (link)]. In order to minimize any air between and within the nanotubes, the solutions were placed in a vacuum (25 in.Hg) chamber for 1 h. Next, the solutions were mixed for 1 h and vacuum was reapplied [14 (link)]. Finally, the HNT+DOX solutions were centrifuged (3000 rpm) for 10 min. The supernatants were collected and stored at −20°C for high-pressure liquid chromatography (HPLC) analysis. Meanwhile, the HNT+DOX pellets were dried at 37°C, weighed, ground, and sieved.

Palasuk J., Windsor L.J., Platt J.A., Lvov Y., Geraldeli S, & Bottino M.C. (2017). Doxycycline-loaded nanotube-modified adhesives inhibit MMP in a dose-dependent fashion. Clinical oral investigations, 22(3), 1243-1252.

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Evaluating Doxycycline and ICG-001 in Liver Cancer Cells

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Doxycycline hyclate (DOX, Sigma); ICG-001 (Selleckchem); fetal bovine serum, DMEM and DMEM/F12 medium (Gibco); fibroblast growth factor (FGF) and epidermal growth factor (EGF) (PeproTech); Trizol reagent (TAKARA); protease and phosphatase inhibitor cocktail (Roche); Lipofectamine 3000 and B27 (Invitrogen); dual-specific luciferase assay kit (Promega); Cell Counting Kit-8 (CCK8, Dojindo Molecular Technologies); Western blotting substrate (Millipore); Cell Signaling Senescence β-Galactosidase Staining Kit (CST); silver staining Rapid silver staining kit's (Beyotime); BALB/c nude mice (Beijing Vital River Laboratory Animal Technology); antibody against GATA4, Lamin B1, P21, FLAG, P15 and c-MYC (Abcam); HA and β-actin (Sigma); β-catenin, LEF1, TCF1, P14, P27, P53, p14/ARF, Caspase-9 and Caspase-3 (CST), P16/Ink4a (Epitomics); Cytokeratin (AE1/AE3) antibody (Kit-0009, MXB Biotechnologies) were purchased from the indicated manufacturers. SNU-387, SNU-449, PLC, NeHepLxHT, SK-Hep1, HepG2, HUH7 and HEK293 cells were obtained from ATCC.

Lu F., Zhou Q., Liu L., Zeng G., Ci W., Liu W., Zhang G., Zhang Z., Wang P., Zhang A., Gao Y., Yu L., He Q, & Chen L. (2020). A tumor suppressor enhancing module orchestrated by GATA4 denotes a therapeutic opportunity for GATA4 deficient HCC patients. Theranostics, 10(2), 484-497.

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Tat Protein Expression Modulation in Transgenic Mice

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Tat protein expression was induced in GT-tg bigenic mice by intraperitoneal administration of Doxycycline hyclate (Dox, Sigma-Aldrich, St. Louis, MO). Dox was prepared fresh daily by dissolving in vehicle (sterile 0.9% saline) to the concentration enabling administration of 0.1 ml per 10 g body weight, and protected from light. Methylsulfonylmethane (MSM, Sigma-Aldrich), which has a half-life in rodents of about 12 hours (84 (link)), was administered intraperitoneally at 100 mg/kg/day, 20 minutes before GT-tg mice were administered saline/Dox, adapted from (85 (link)). Diethyl maleate (DEM, Sigma-Aldrich), which rapidly depletes GSH with a half-life of about 15 hours (64 (link)), was given intraperitoneally for one or two days. DEM was administered at a dose of 34.8 μM in 0.25 ml saline, and in pretreatment studies was given 30 minutes before saline or Dox based on prior studies in mice (65 (link),86 (link)).

McLaughlin J.P., Paris J.J., Mintzopoulos D., Hymel K.A., Kim J.K., Cirino T.J., Gillis T.E., Eans S.O., Vitaliano G.D., Medina J.M., Krapf R.C., Stacy H.M, & Kaufman M.J. (2017). Conditional Human Immunodeficiency Virus Transactivator of Transcription Protein Expression Induces Depression-like Effects and Oxidative Stress. Biological psychiatry. Cognitive neuroscience and neuroimaging, 2(7), 599-609.

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Purification and Labeling of Antibodies

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Doxycycline hyclate (Dox) was purchased from Sigma-Aldrich. Nickel-nitrilotriacetic acid (NTA) beads were purchased from Qiagen. Mouse anti-HA.11 (clone 16B12) coupled to Alexa Fluor (AF) 488 was purchased from Life Technologies. Mouse anti-HA.11 (clone 16B12) was acquired from BioLegend. VHHs coupled to either Alexa Fluor (AF) 647 or 5-carboxytetramethylrhodamine (TAMRA) were generated using sortase A as described earlier (44 (link)). Hybridoma cells secreting mouse monoclonal anti-IAV NP (clone H16-L10-4R5; ATCC HB-65) were obtained from ATCC, and antibodies in the supernatant were purified using a protein G column.

Hanke L., Knockenhauer K.E., Brewer R.C., van Diest E., Schmidt F.I., Schwartz T.U, & Ploegh H.L. (2016). The Antiviral Mechanism of an Influenza A Virus Nucleoprotein-Specific Single-Domain Antibody Fragment. mBio, 7(6), e01569-16.

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Antibody-Based Protein Detection Protocol

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Antibodies against β-Actin, Bcl2, p-ERK, ERK, LC3, p62, XIAP, ATG5 and PARP were purchased from Cell Signaling Technology (Danvers, MA, USA). Anti-XIAP and anti-pP27 were purchased from BD Biosciences (San Jose, CA, USA). Anti-Caspase-3 (Active) antibody was purchased from Millipore (Temecula, CA, USA). An anti-ARHI murine monoclonal antibody (ID8) was generated in our laboratory. Doxycycline hyclate (DOX), chloroquine diphosphate salt (CQ), cis-diammine-platinum (II) dichloride (Cisplatin), Z-VAD, Nec-1 and N-acetyl-L-cysteine were purchased from Sigma-Aldrich (St. Louis, MO, USA).

Washington M.N., Suh G., Orozco A.F., Sutton M.N., Yang H., Wang Y., Mao W., Millward S., Ornelas A., Atkinson N., Liao W., Bast RC J.r, & Lu Z. (2015). ARHI (DIRAS3)-mediated autophagy-associated cell death enhances chemosensitivity to cisplatin in ovarian cancer cell lines and xenografts. Cell Death & Disease, 6(8), e1836-.

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Doxycycline-Induced HIV-1 Tat Expression

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All protocols followed the guidelines for the use of laboratory animals and were approved by the Institutional Animal Care and Usage Committee at Temple University.
This study used brain tissue of doxycycline induced GFAP promoter-driven HIV-1 Tat transgenic mice generously provided by Dr. Johnny He to the Comprehensive NeuroAIDS Center^{47 (link)}. Briefly, mice were divided into two groups, (+DOX) and (−DOX) with 4 animals per group. The doxycycline-induced Tat (iTat) animals were injected i.p. with doxycycline hyclate (DOX) (Sigma-Aldrich) at the dosage of 80 mg/kg/day for seven days. The control group was injected with saline (.09% NaCl) in the same manner as their Dox treated counterparts. After seven days, both groups of mice were sacrificed and brains were extracted/dissected into regions of interest (i.e. hippocampus, cerebellum, brain stem, and frontal cortex)^{55 (link)}. Total RNA was isolated for each brain region using Trizole reagent (Invitrogen, ThermoFisher, Carlsbad, CA), as previously described^{55 (link)}.

Torkzaban B., Natarajaseenivasan K., Mohseni Ahooyi T., Shekarabi M., Amini S., Langford T.D, & Khalili K. (2020). The lncRNA LOC102549805 (U1) modulates neurotoxicity of HIV-1 Tat protein. Cell Death & Disease, 11(10), 835.

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Inducible Epithelial-Mesenchymal Transition in LNCaP Cells

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LNCaP (ATCC), Rockville, MD), LNCaP–iSnail, LNCaP–iSlug, and –iGFP cells were grown in phenol red free Roswell Park Memorial Institute—1640 medium (RPMI) (Thermofisher) containing 5% foetal bovine serum (FBS; ThermoFisher), 1% streptomycin –penicillin, and 0.05% gentamycin (Life Technologies) at 37 °C with 5% CO₂. HEK293T (ATCC) cultured in Dulbecco’s modified Eagle’s medium (DMEM) supplemented with 10% heat inactivated FBS at 37 °C with 5% CO₂. The pENTR223.1 entry clone containing the cDNA of SNAI1 (Snail) and eGFP were recombined into the pINDUCER20 lentiviral construct [13 (link)] using Gateway^® LR Clonase^® II enzyme mix according to manufacturer’s instructions (Thermofisher). Lentiviral packaging and supernatants were generated by transient transfection of HEK293T cells (ATCC) as described previously [67 (link)]. Stable LNCaP (ATCC) sublines expressing pINDUCER20-Snail, pINDUCER20-Slug, or pINDUCER20-GFP were generated by lentiviral transduction followed by selection with 1 mg/mL G418 (Invivogen). Optimal doxycycline hyclate (Dox; Sigma Aldrich) concentration for induction of cDNA expression was determined to be 1 μg/mL (data not shown) and was refreshed every 48 h of cell culture.

Stylianou N., Lehman M.L., Wang C., Fard A.T., Rockstroh A., Fazli L., Jovanovic L., Ward M., Sadowski M.C., Kashyap A.S., Buttyan R., Gleave M.E., Westbrook T.F., Williams E.D., Gunter J.H., Nelson C.C, & Hollier B.G. (2018). A molecular portrait of epithelial–mesenchymal plasticity in prostate cancer associated with clinical outcome. Oncogene, 38(7), 913-934.

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