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D9891

Manufactured by Merck Group
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D9891 is a laboratory equipment product manufactured by Merck Group. It is designed to perform specific functions in a laboratory setting. The core function of this product is to facilitate accurate measurements and data collection for research and analysis purposes. No further details about its intended use or application are provided.

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Lab products found in correlation

Doxycycline, by Merck Group (7 mentions) Fetal bovine serum (fbs), by Thermo Fisher Scientific (3 mentions) Penicillin streptomycin, by Thermo Fisher Scientific (3 mentions) M0521, by Merck Group (2 mentions) E10521, by Merck Group (2 mentions) P8833, by Merck Group (2 mentions) Tet plko puro plasmid, by Addgene (2 mentions) Puromycin, by Thermo Fisher Scientific (2 mentions) Polyethylenimine (pei), by Merck Group (2 mentions) Polybrene, by Merck Group (2 mentions) E8875, by Merck Group (1 mentions) Bp2818 4, by Thermo Fisher Scientific (1 mentions) Genetecin, by Thermo Fisher Scientific (1 mentions) Opti mem, by Thermo Fisher Scientific (1 mentions) Puromycin, by Merck Group (1 mentions) S3888, by Bio-Serv (1 mentions) Tamoxifen, by Merck Group (1 mentions) Hepes, by Thermo Fisher Scientific (1 mentions) Charcoal stripped fetal bovine serum, by Merck Group (1 mentions) Lipofectamine 3000 reagent, by Thermo Fisher Scientific (1 mentions)

Spelling variants of this product

Doxycycline (D9891) (10 citations) Doxycycline hyclate (D9891) (5 citations)

39 protocols using d9891

1

Yeast Culture Compound Screening

Cited 2 times
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The following molecules were incubated in yeast cultures for 10 min at the following concentrations. Rapamycin (TORC1 inhibitor, 200 ng/mL - Fisher Scientific BP2963-1), dissolved in DMSO (Amresco 0231); 1,10 Phenanthroline (RNAP I–III inhibitor, 100 µg/mL - Sigma-Aldrich 131377), in ethanol (Fisher Scientific BP28184); doxycycline (Antibiotic/regulator of Tet-Off/On promoters; 10 µg/mL - Sigma-Aldrich D-9891) in water, β-estradiol (regulator of β-estradiol promoter; 1 µM - Sigma-Aldrich E8875), in ethanol; thiolutin (RNAP I-III inhibitor, 3 µg/mL - VWR 89157-260), in DMSO; 4tU (Uracil analog, 25 µg/mL - Sigma-Aldrich 440736), in DMSO. All drug concentrations are based on standards in the field and/or the following studies (Grigull et al. 2004 (link); Mnaimneh et al. 2004 (link); Munchel et al. 2011 (link)). Ten minutes was chosen as a time period for analysis given that mRNA half-lives were recently reported to average 5 min (Chan et al. 2018 (link)), thus short-term effects are relevant for accurate half-life measurement.
Eshleman N., Luo X., Capaldi A, & Buchan J.R. (2020). Alterations of signaling pathways in response to chemical perturbations used to measure mRNA decay rates in yeast. RNA, 26(1), 10-18.
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2

Doxycycline-Inducible BRN2 Flag Cell Line Generation

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The FuGENE 6 lipofection system was used to generate polyclonal stable, doxycycline-inducible BRN2 Flag cell lines by cotransfecting PB Brn2-Flag wild-type or truncation mutants and were cotransfected with the PB transposase vector pPyCAG-PBase and the Tet-On System vector with a Neomycin resistance cassette pPB-CAG-rtTA-IRES-Neo. Transfection reagents were prepared in OptiMEM (Gibco, Thermo Fisher) at a FuGENE to DNA plasmid ratio of 3:1. Following transfection, cell lines were subjected to double selection with 3 µg/mL puromycin (Sigma-Aldrich, p8833) and 750 µg/mL Genetecin (Gibco, ThermoFisher, G418) for 48–72 h until the death of all control 501mel cells treated in parallel. Monoclonal cell lines from BRN2-Flag wild type were established by colony picking and were screened by immunofluorescence. Cells were induced with doxycycline as indicated (Sigma-Aldrich, D9891).
Herbert K., Binet R., Lambert J.P., Louphrasitthiphol P., Kalkavan H., Sesma-Sanz L., Robles-Espinoza C.D., Sarkar S., Suer E., Andrews S., Chauhan J., Roberts N.D., Middleton M.R., Gingras A.C., Masson J.Y., Larue L., Falletta P, & Goding C.R. (2019). BRN2 suppresses apoptosis, reprograms DNA damage repair, and is associated with a high somatic mutation burden in melanoma. Genes & Development, 33(5-6), 310-332.
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3

shRNA-mediated α-catenin knockdown

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siRNA sequences targeting mouse (5'‐GTCACATGCTTCACTCAAA‐3') and human (5'‐GTCACTGTTCGTCACTCAA‐3') α‐catenin were cloned as shRNA in the lentiviral vector pFUTG as described previously 7. The shRNA against mouse α‐catenin was positioned in the 5'‐UTR. Lentiviral particles were produced in COS‐7 cells and used for transduction as described previously 7. To induce knockdown, cells were treated with 2 mg/ml doxycycline (1:5000, D9891; Sigma‐Aldrich, Zwijndrecht, The Netherlands) for at least 3 days.
de Groot J.S., Ratze M.A., van Amersfoort M., Eisemann T., Vlug E.J., Niklaas M.T., Chin S., Caldas C., van Diest P.J., Jonkers J., de Rooij J, & Derksen P.W. (2018). αE‐catenin is a candidate tumor suppressor for the development of E‐cadherin‐expressing lobular‐type breast cancer. The Journal of Pathology, 245(4), 456-467.
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4

Sciatic Nerve Regeneration in Rats

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The sciatic nerve defect model was established in 30 male Wistar rats (150–180 g, 8-weeks-old) and LvmiR-338-mCherry was injected in nerve conduits. The animals were randomized into two groups (15 rats/group); one group was treated with doxycycline and the control group received sucrose. From previous experiments, treatments (doxycycline (200 ng/mL), Sigma, D9891; and sucrose (1%) in drinking water) were started at the 3rd day after surgery. The distal sciatic nerves were collected in 1, 3, 5, 7 and 9 days after doxycycline treatment (n = 3 rats/time point).
The distal nerve sections in each group were examined by fluorescence observation as described above.
Wen L.L., Yu T.H., Ma Y.Z., Mao X.Y., Ao T.R., Javed R., Ten H., Matsuno A, & Ao Q. (2022). Sequential expression of miR-221-3p and miR-338-3p in Schwann cells as a therapeutic strategy to promote nerve regeneration and functional recovery. Neural Regeneration Research, 18(3), 671-682.
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5

Doxycycline-induced Autophagy Inhibition

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Blood glucose measurements were taken prior to, and after 24 h of, food withdrawal using the ACCU-CHEK Aviva blood glucose monitor. As doxycycline is provided in the diet of mice (Test Diets, 5A5X), mice also received doxycycline intraperitoneally (20 ml/kg of a 4 mg/ml solution, Sigma, D9891) to ensure continued expression of shAtg5.
Cassidy L.D., Young A.R., Pérez-Mancera P.A., Nimmervoll B., Jaulim A., Chen H.C., McIntyre D.J., Brais R., Ricketts T., Pacey S., De La Roche M., Gilbertson R.J., Rubinsztein D.C, & Narita M. (2018). A novel Atg5-shRNA mouse model enables temporal control of Autophagy in vivo. Autophagy, 14(7), 1256-1266.
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6

Conditional Gene Recombination in Mice

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For TetO-Cre–mediated gene recombination, mice harboring M2-rtTA and TetO-Cre as well as targeted Gfi-1b alleles (age 4–8 wk) were subjected to daily doxycycline injections (D9891; Sigma-Aldrich; 10 µg/g mouse weight; diluted 1 mg/ml in PBS) and doxycycline-supplemented drinking water (2 mg/ml doxycycline, 10 mg/ml sucrose) for up to 4 wk. For Mx-Cre–mediated gene recombination, a total of three intraperitoneal injections (at 8.5 µg/g mouse bodyweight) of pIpC (GE Healthcare) were administered every other day.
Foudi A., Kramer D.J., Qin J., Ye D., Behlich A.S., Mordecai S., Preffer F.I., Amzallag A., Ramaswamy S., Hochedlinger K., Orkin S.H, & Hock H. (2014). Distinct, strict requirements for Gfi-1b in adult bone marrow red cell and platelet generation. The Journal of Experimental Medicine, 211(5), 909-927.
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7

Activation of Gene Expression in Mice

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Substances were administered to genetically modified mice, in order to activate gene expression and label dividing cells for in vivo experiments only. This includes doxycycline food (200 mg/kg of chow, BioServ #S3888), which replaced the dam’s normal chow. Additionally, three substances were administered via injections to post-natal day 0 to post-natal day 2 (P0-P2) pups intramuscularly, in the upper thigh, using Ultrafine insulin syringes (Becton-Dickinson 31G #08290–328468). These included doxycycline hyclate (DOX: 100 mg/kg body weight, Sigma Aldrich #D9891), which was freshly prepared as 10 mg/ml stock in 0.9% sterile saline. 5-ethynyl-2´-deoxyuridine (EdU: 0.01 mg/kg, Invitrogen #A10044) was made as a 10 mM stock solution in DMSO and diluted for injection to 40% strength in 0.9% sterile saline. Tamoxifen (75 mg/kg, Sigma, #T5648) was dissolved in corn oil (Sigma, #C8267) at 5 mg/kg.
Zhang J., Wang Q., Abdul-Aziz D., Mattiacio J., Edge A.S, & White P.M. (2018). ERBB2 signaling drives supporting cell proliferation in vitro and apparent supernumerary hair cell formation in vivo in the neonatal mouse cochlea. The European journal of neuroscience, 48(10), 3299-3316.
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8

Xenotransplantation of AML in NSG Mice

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Adult NOD.Cg-PrkdcscidIl2rgtm1Wjl/SzJ (NSG) mice (6–8 weeks old) were pretreated with 280 cGy total body irradiation, then 5 × 105 viable OCI-AML3 cells (in 300 μl PBS) or 1 × 106 patient BM cells (in 300 μl PBS) were injected into tail veins of the recipient NSG mice for transplantation. After transplantation, the recipients mice were administered with (for OCI-AML3 cells stably expressing inducible shHOXBLINC to induce HoxBlinc KD) or without (for AML cells with dCas9-mediated HoxBlinc inactivation) doxycycline in the drinking water (Sigma D-9891, 1 mg/ml, 1% sucrose, newly prepared every other day) until being sacrificed, and daily monitored for symptoms (ruffled coat, hunched back, weakness, and reduced motility) and survival time. For each set of xenotransplantation, recipient mice of all groups were killed and analyzed on the same day when any group of recipients exhibited a moribund condition. Human CD45 chimerism in the BM, spleen cells, and PB WBC were analyzed by flow cytometry, histological assay, IHC for hCD45 were also performed as described above.
Zhu G., Luo H., Feng Y., Guryanova O.A., Xu J., Chen S., Lai Q., Sharma A., Xu B., Zhao Z., Feng R., Ni H., Claxton D., Guo Y., Mesa R.A., Qiu Y., Yang F.C., Li W., Nimer S.D., Huang S, & Xu M. (2021). HOXBLINC long non-coding RNA activation promotes leukemogenesis in NPM1-mutant acute myeloid leukemia. Nature Communications, 12, 1956.
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9

Merlin Expression and Radiation Therapy in Meningioma Xenografts

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This study was approved by the UCSF Institutional Animal Care and Use Committee (AN174769), and all experiments complied with relevant ethical regulations. Xenograft experiments were performed by implanting 3 million CH-157MN cells into the flank of 5–6-week-old female NU/NU mice (Harlan Sprague Dawley Inc.). To induce Merlin expression in meningiomas in vivo, mice harboring CH-157MN cells encoding pLV.APEX2-Merlin were treated with doxycycline 200 μg/ml (n=3) (#D9891, Sigma) or vehicle (n=3) 14 days post implantation. After 7 days of treatment, 2 Gy of ionizing radiation per day was delivered using a Precision X-RAD 320 Cabinet Irradiation, with normal operating settings, on each of 2 successive days. Tumors were collected 24 hours after the second dose of ionizing radiation for immunoblotting and immunohistochemistry. For preclinical pharmacologic experiments, animals in the treatment arm (n=7) were gavaged with 100 μg/g abemaciclib in 0.5% methylcellulose vehicle daily starting 12 days after injection, until protocol-defined endpoints. Animals in the vehicle arm (n=13) were gavaged at the same frequency for the same duration with 0.5% methylcellulose. For Kaplan-Meier survival analysis, events were recorded when tumors reached the protocol-defined size of 2000 mm3.
Choudhury A., Magill S.T., Eaton C.D., Prager B.C., Chen W.C., Cady M.A., Seo K., Lucas C.H., Casey-Clyde T.J., Vasudevan H.N., Liu S.J., Villanueva-Meyer J.E., Lam T.C., Pu J.K., Li L.F., Leung G.K., Swaney D.L., Zhang M.Y., Chan J.W., Qiu Z., Martin M.V., Susko M.S., Braunstein S.E., Bush N.A., Schulte J., Butowski N., Sneed P.K., Berger M.S., Krogan N.J., Perry A., Phillips J.J., Solomon D.A., Costello J.F., McDermott M.W., Rich J.N, & Raleigh D.R. (2022). Meningioma DNA methylation groups identify biological drivers and therapeutic vulnerabilities. Nature genetics, 54(5), 649-659.
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10

Doxycycline-Induced Liver Transformation

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Induction of transgene expression were conducted in 3-month-old adult fish for 7 days with 60 μg/ml doxycycline (D9891; Sigma). In preliminary experiments, we found no histological change of liver histology after one day of doxycycline induction but histological transformation of liver cells (hyperplasia) was observed from 3 days post-induction (data not shown). At the end of doxycycline treatment, >10 fish in each group were used for imaging analyses. All the zebrafish were anesthetized in 0.08% tricaine (E10521; Sigma) and immobilized in 3% methylcellulose (M0521; Sigma) before imaging. Each fish was photographed individually from the left lateral side with an Olympus microscope.
Yang Q., Yan C, & Gong Z. (2017). Activation of liver stromal cells is associated with male-biased liver tumor initiation in xmrk and Myc transgenic zebrafish. Scientific Reports, 7, 10315.
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