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Dmpge2

Manufactured by Cayman Chemical
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DmPGE2 is a synthetic analog of prostaglandin E2 (PGE2). It is a chemical compound used in research and laboratory applications.

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Lab products found in correlation

Ns 398, by Cayman Chemical (3 mentions) Poly 1 c hmw, by InvivoGen (2 mentions) G csf recombinant human protein, by Thermo Fisher Scientific (2 mentions) Indomethacin, by Merck Group (2 mentions) Iccb known bioactive library, by Enzo Life Sciences (2 mentions) As605240, by Merck Group (2 mentions) 11 12 eet, by Cayman Chemical (2 mentions) P3 primary cell 4d nucleofector x kit, by Lonza (2 mentions) Flt3l, by STEMCELL (2 mentions) Stemspan, by STEMCELL (2 mentions) Methocult h4434, by STEMCELL (2 mentions) Sfem 2 medium, by STEMCELL (2 mentions) Human cord blood cd34 positive selection kit 2, by STEMCELL (2 mentions) Clinmacs cd34 microbead kit, by Miltenyi Biotec (2 mentions) Um171, by STEMCELL (2 mentions) Cd34 expansion supplement, by STEMCELL (2 mentions) Poloxamer 407, by BASF (2 mentions) Penicillin, by Thermo Fisher Scientific (2 mentions) Streptomycin, by Thermo Fisher Scientific (2 mentions) L 902 688, by Cayman Chemical (1 mentions)

21 protocols using dmpge2

1

Therapeutic Interventions for Radiation-Induced Injury

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Mice in the dmPGE2-treated groups received single intraperitoneal (IP) injections of either 2 mg/kg or 6 mg/kg body weight dmPGE2 (Cayman Chemical, Ann Arbor, MI) dissolved in methyl acetate at 24 h after TBI in the lethal and sublethal models, respectively. Irradiated control mice received a single IP injection of methyl acetate (Sigma-Aldrich® LLC, St. Louis, MO) vehicle at the same dosage and timing as the dmPGE2-treated groups. All mice also received 500 μl (60 mg/l in the drinking water; Cayman Chemicals; estimated to provide a dose of 28.5 mg/m2/day), was initiated at day 7 after TBI in the lethal model. The dose and schedule of lisinopril are equivalent and consistent with previously published studies in a rat model (43 (link)); however, water consumption was not quantitated to account for variation in lisinopril dosage, particularly after irradiation. Lisinopril was started at day 3 postirradiation in the sublethal model to begin therapy more than 24 h after TBI but prior to the decline in peripheral platelet counts (45 (link)). All lisinopril-treated groups continued therapy for the duration of the study and water packs containing lisinopril were replaced weekly.
Saunders J I.I., Niswander L.M., McGrath K.E., Koniski A., Catherman S.C., Ture S.K., Medhora M., Kingsley P.D., Calvi L.M., Williams J.P., Morrell C.N, & Palis J. (2021). Long-acting PGE2 and Lisinopril Mitigate H-ARS. Radiation research, 196(3), 284-296.
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2

Indomethacin Modulation of Hematopoietic Stem Cells

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For the HSC Replicate 1 experiment, we used the following mouse strain (#016617) that was obtained from Jackson labs but bred in-house. For external stimulant treatments, male and female mice (8–10 weeks) were ordered from Jackson labs (strain CD 45.2 [Ly5.2], #00664). Mice were kept for at least 1 week in the animal facility before initiating experiments and allocated at random (by cage) into experimental groups. Indomethacin (Sigma, 6 mg/l) was administered for 7 days in acidified drinking water to maintain stability (Curry et al., 1982 (link); Praticò et al., 2001 (link)). Indomethacin supplemented drinking water was changed every other day. Mice were injected with the following drugs and euthanized after 2 hr: poly(I:C) HMW (Invivogen), IP injection 10 mg/kg (Pietras et al., 2014 (link)). G-CSF Recombinant Human Protein (Thermo Fisher), IP injection, 0.25 mg/kg (Morrison et al., 1997 (link)). dmPGE2 (Cayman), SC injection, 2 mg/kg (Hoggatt et al., 2013 (link)). Mice were weighed before injection and injection volume was adjusted to ensure equal dose between individual mice. The ‘control’ condition from the external stimulant treatments was also used as the second independent biological replicate of unperturbed HSCs (HSC Replicate 2). All animal procedures were approved by the Harvard University Institutional Animal Care and Use Committee.
Fast E.M., Sporrij A., Manning M., Rocha E.L., Yang S., Zhou Y., Guo J., Baryawno N., Barkas N., Scadden D., Camargo F, & Zon L.I. (2021). External signals regulate continuous transcriptional states in hematopoietic stem cells. eLife, 10, e66512.
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3

Agonists for Radioprotective Effects

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DmPGE2, the EP1,3 dual agonist 17-phenyl trinor PGE2, the EP2 selective agonist Butaprost (free acid), and the selective EP4 agonist L-902,688, were purchased from Cayman Chemical (Ann Arbor, MI), and were used as described elsewhere (24 (link), 25 (link)). DmPGE2, and Butaprost in MeAc and L-902,688 in MeOH were evaporated under a stream of N2 and reconstituted in absolute EtOH. Crystalline 17-phenyl trinor PGE2 was solubilized in absolute EtOH. All compounds were diluted in phosphate buffered saline (PBS) for injection. DmPGE2 was injected as a single subcutaneous (s.c.) dose of 35 μg (containing 1.75% EtOH) or two doses of 20 μg (containing 1% EtOH) per mouse at different times relative to irradiation as indicated. The range of doses of dmPGE2 as a function of all mice used was 1.73 mg/kg for pediatric, 1.6 mg/kg for young adult, and 0.91 mg/kg for geriatric mice, a <2-fold variation among all sexes and ages of mice. EP receptor agonists were injected 30 min prior to TBI as a single s.c. dose of 35 μg (containing 1.75% EtOH) or 10 μg (containing 0.5% EtOH) per mouse. All compounds were injected in a volume of 200 μl per mouse. In all cases, vehicle injections contained equivalent EtOH concentrations.
Patterson A.M., Wu T., Chua H.L., Sampson C.H., Fisher A., Singh P., Guise T.A., Feng H., Muldoon J., Wright L., Plett P.A., Pelus L.M, & Orschell C.M. (2021). Optimizing and Profiling Prostaglandin E2 as a Medical Countermeasure for the Hematopoietic Acute Radiation Syndrome. Radiation research, 195(2), 115-127.
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4

Screening Zebrafish Immune Cells

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A total of 200,000 WKM cells was incubated with zebrafish medium containing 50 μM dmPGE2 (Cayman Chemical) in DMSO for 2 hr on ice. DMSO was used as vehicle control. Cells were then washed twice with and suspended in PBS before delivery to recipient fish as described before. The chemical screen was performed using the NatProd Collection from MicroSource Discovery Systems. All chemicals were in DMSO at a concentration of 10 mM and diluted to 10 μM when mixed with donor WKM cells. Transplants were performed as stated above.
Astuti Y., Kramer A.C., Blake A.L., Blazar B.R., Tolar J., Taisto M.E, & Lund T.C. (2016). A Functional Bioluminescent Zebrafish Screen for Enhancing Hematopoietic Cell Homing. Stem Cell Reports, 8(1), 177-190.
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5

Chemical Screen for Zebrafish Stem Cell

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The ICCB Known Bioactive Library was purchased from BIOMOL (Enzo Life Sciences) and used for the adult zebrafish transplantation-based chemical screen. Chemicals were diluted at a 1:200 ratio. Chemicals used for the secondary round of screening for confirmation were from a different aliquot of the library, independent of the primary screen plate. 11,12-EET (Cayman Chemical, Cat. 50511) was resuspended in DMSO with original organic solvent evaporated. AS605240 (Sigma-Aldrich Cat. A0233) was resuspended in DMSO. The following chemicals were used for zebrafish marrow treatment: dmPGE2 (Cayman, Cat. 14750), 10 μM; BIO (EMD), 0.5 μM. 0.5 μM 11,12-EET and 14,15-EET were used for zebrafish WKM treatment (Fig. 1e); 2 μM 11,12-EET for all mouse WBM treatment (Fig. 4); 5 μM 11,12-EET for all zebrafish embryo treatment (Fig. 2, 3). The concentrations were chosen based on dose titration pilot experiments with doses spanning 0.1–50 μM. For the chemical suppressor screen, the suppressors were added 30 min prior to 11,12-EET. Zebrafish embryos were incubated with inhibitors at three different concentrations. The highest effective concentrations tested without causing general toxicity are listed in Supplementary Table S1.
Li P., Lahvic J.L., Binder V., Pugach E.K., Riley E.B., Tamplin O.J., Panigrahy D., Bowman T.V., Barrett F.G., Heffner G.C., McKinney-Freeman S., Schlaeger T.M., Daley G.Q., Zeldin D.C, & Zon L.I. (2015). Epoxyeicosatrienoic Acids Enhance Embryonic Haematopoiesis and Adult Marrow Engraftment. Nature, 523(7561), 468-471.
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6

CD34+ Cell Expansion and Genetic Modification

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CD34+ cells were either freshly purified from human CB after obtaining informed consent and upon approval by the San Raffaele Hospital Bioethical Committee, or purchased frozen from Lonza. 106 CD34+ cells/ml were stimulated in serum-free StemSpan medium (StemCell Technologies) supplemented with penicillin, streptomycin and human early-acting cytokines (for CB-derived cells: stem cell factor (SCF) 100 ng/ml, Flt3 ligand (Flt3-L) 100 ng/ml, thrombopoietin (TPO) 20 ng/ml, and interleukin 6 (IL-6) 20 ng/ml; for BM-derived cells: SCF 300ng/ml, Flt3-L 300 ng/ml, TPO 100 ng/ml, and IL-6 60 ng/ml; all purchased from Peprotech) for 24 or 48 hr and then infected with IDLVs at multiplicity of infection (MOI) 100-500. The following day the cells were electroporated with 175 μg/ml ZFNs encoding mRNAs (P3 Primary Cell 4D-Nucleofector X Kit, program EO-100; Lonza). For some experiments, the following drugs were supplemented to the culture media: 1 μM SR1 (kindly provided by T. Boitano and M. Cooke, GNF) added at every medium change, and 10 μM dmPGE2 (Cayman) added at the beginning of the culture, 1 hour before and just after electroporation. For CFC assays, 800 cells/plate were seeded one day after electroporation in methylcellulose-based medium (MethoCult H4434, StemCell Technologies). Two weeks after plating, colonies were counted and identified according to morphological criteria.
Genovese P., Schiroli G., Escobar G., Tomaso T.D., Firrito C., Calabria A., Moi D., Mazzieri R., Bonini C., Holmes M.C., Gregory P.D., van der Burg M., Gentner B., Montini E., Lombardo A, & Naldini L. (2014). Targeted Genome Editing in Human Repopulating Hematopoietic Stem Cells. Nature, 510(7504), 235-240.
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7

CD34+ Cell Expansion and Genetic Modification

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CD34+ cells were either freshly purified from human CB after obtaining informed consent and upon approval by the San Raffaele Hospital Bioethical Committee, or purchased frozen from Lonza. 106 CD34+ cells/ml were stimulated in serum-free StemSpan medium (StemCell Technologies) supplemented with penicillin, streptomycin and human early-acting cytokines (for CB-derived cells: stem cell factor (SCF) 100 ng/ml, Flt3 ligand (Flt3-L) 100 ng/ml, thrombopoietin (TPO) 20 ng/ml, and interleukin 6 (IL-6) 20 ng/ml; for BM-derived cells: SCF 300ng/ml, Flt3-L 300 ng/ml, TPO 100 ng/ml, and IL-6 60 ng/ml; all purchased from Peprotech) for 24 or 48 hr and then infected with IDLVs at multiplicity of infection (MOI) 100-500. The following day the cells were electroporated with 175 μg/ml ZFNs encoding mRNAs (P3 Primary Cell 4D-Nucleofector X Kit, program EO-100; Lonza). For some experiments, the following drugs were supplemented to the culture media: 1 μM SR1 (kindly provided by T. Boitano and M. Cooke, GNF) added at every medium change, and 10 μM dmPGE2 (Cayman) added at the beginning of the culture, 1 hour before and just after electroporation. For CFC assays, 800 cells/plate were seeded one day after electroporation in methylcellulose-based medium (MethoCult H4434, StemCell Technologies). Two weeks after plating, colonies were counted and identified according to morphological criteria.
Genovese P., Schiroli G., Escobar G., Tomaso T.D., Firrito C., Calabria A., Moi D., Mazzieri R., Bonini C., Holmes M.C., Gregory P.D., van der Burg M., Gentner B., Montini E., Lombardo A, & Naldini L. (2014). Targeted Genome Editing in Human Repopulating Hematopoietic Stem Cells. Nature, 510(7504), 235-240.
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8

dmPGE2 Delivery via Mini-Osmotic Pump

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Mice were treated with dmPGE2 (Cayman Chemical) and vehicle control through a subcutaneously implanted ALZET 1004 mini–osmotic pump (DURECT). Minipumps were loaded with 100 μL of dmPGE2 solution in sterile PBS. A release rate of 0.11 μL/h administered a total amount of 30 μg/kg dmPGE2 daily, a dose that resulted in elevated serum PGE2 without obvious adverse side effects, during the 28 days of the experimental setup.
Liu D., Ding Q., Dai D.F., Padhy B., Nayak M.K., Li C., Purvis M., Jin H., Shu C., Chauhan A.K., Huang C.L, & Attanasio M. (2021). Loss of diacylglycerol kinase ε causes thrombotic microangiopathy by impairing endothelial VEGFA signaling. JCI Insight, 6(9), e146959.
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9

Maternal Exposure to dmPGE2 in Mice

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Male and female mice were mated overnight, and the females were checked every morning until a vaginal plug was observed. This day was considered as gestation day 1, and the females were housed separately for the remaining time. On gestation day 11, the pregnant females were weighed, and injected subcutaneously with 0.2 mg/kg concentration of 16,16-dimethyl prostaglandin E2 (dmPGE2; Cayman Chemical) in saline as in previous studies [18] (link), [65] (link). The compound dmPGE2 was used as it has a slower metabolism rate than PGE2, and therefore remains active for a longer period of time [44] (link), [58] (link). Control animals were injected with saline only. Three pregnant females were administered with a single dose of saline or the PGE2 compound, resulting in three litters for each developmental stage tested. Embryonic day 11 (E11) was chosen as the day of PGE2 exposure to the embryo as it is the onset of neurogenesis in the embryonic mouse brain, and also equivalent to the time during which the analogous drug misoprostol was consumed in human clinical studies that resulted in manifestation of Moebius syndrome and ASD [45] (link), [7] (link).
Rai-Bhogal R., Wong C., Kissoondoyal A., Davidson J., Li H, & Crawford D.A. (2018). Maternal exposure to prostaglandin E2 modifies expression of Wnt genes in mouse brain – An autism connection. Biochemistry and Biophysics Reports, 14, 43-53.
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10

Modulation of Wound Healing in Zebrafish

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Larvae were treated with 30 μM NS398 or 20 μM dmPGE2 (both from Cayman Chemicals) immediately after laser wounding at 2 or 3 dpf in Danieau’s solution containing 0.5% DMSO. After treatment, larvae were fixed in 4% PFA overnight, immunostained and imaged as described above. For DPI treatment, larvae were incubated in 100 μM DPI (Sigma) in Danieau’s solution containing 1% DMSO for 45 min prior to wounding and throughout the period of imaging.
Antonio N., Bønnelykke-Behrndtz M.L., Ward L.C., Collin J., Christensen I.J., Steiniche T., Schmidt H., Feng Y, & Martin P. (2015). The wound inflammatory response exacerbates growth of pre-neoplastic cells and progression to cancer. The EMBO Journal, 34(17), 2219-2236.
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