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Polyinosinic polycytidylic acid pipc

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Polyinosinic-polycytidylic acid (pIpC) is a synthetic double-stranded RNA molecule used as a laboratory reagent. It functions as a Toll-like receptor 3 (TLR3) agonist, activating the innate immune response.

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Lab products found in correlation

Fetal bovine serum (fbs), by Thermo Fisher Scientific (3 mentions) Rosa26 eyfp, by Jackson ImmunoResearch (2 mentions) Lck cre 003802, by Jackson ImmunoResearch (2 mentions) Cd8a cre 008766, by Jackson ImmunoResearch (2 mentions) Rosa26nicd, by Jackson ImmunoResearch (2 mentions) Penicillin streptomycin, by Thermo Fisher Scientific (2 mentions) Vav cre, by Jackson ImmunoResearch (1 mentions) Mx cre, by Jackson ImmunoResearch (1 mentions) Mx1 cre mice, by Shanghai Model Organisms (1 mentions) Tamoxifen, by Merck Group (1 mentions) Mmtv cre, by Jackson ImmunoResearch (1 mentions) Tg mmtv cre 4mam j, by Jackson ImmunoResearch (1 mentions) Polybrene transfection reagent, by Merck Group (1 mentions) Protease inhibitor cocktail, by Nacalai Tesque (1 mentions) Anti h3k27ac, by Merck Group (1 mentions) Human m csf, by Thermo Fisher Scientific (1 mentions) Lipofectamine 2000, by Thermo Fisher Scientific (1 mentions) Anti h3k27me3, by Merck Group (1 mentions) Anti h3k4me3 antibody, by Active Motif (1 mentions)

16 protocols using polyinosinic polycytidylic acid pipc

1

Conditional hRasGRP1 Expression Model

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The expression construct was generated by cloning the various components into targeting ROSA vector (ROSA-HR). Vector was obtained by introducing first a flanked Neo/STOP cassette and then the hRasGRP1 cDNA cassette, IRES-EGFP, and human polA sequences into a Rosa26 targeting vector that already contain a ubiquitous CAG promoter cassette (Figure. 1A). This construct was transfected into ES cells by electroporation in optimized condition (5×106 cells in presence of 40 μg linearized plasmid, 260V, 500 uF). Positive selection started 48 h after electroporation by adding 200 μg/ml of G418 in 96-well plates. Positive clones were further screened by southern blotting. Furthermore, the hybridization with internal probe did not indicate that any of the ES cells screened contained an additional randomly integrated copy of the targeting plasmid. At the age of 4 weeks mice received a single intraperitoneal injection of polyinosinic-polycytidylic acid (pIpC) (1μg/250μl/mouse) (Sigma, San Luis, MO, US) to induce the expression of hRASGRP1 and GFP from the Rosa26 locus.
Karra L., Romero-Moya D., Ksionda O., Krush M., Gu Z., Mues M., Depeille P., Mullighan C, & Roose J.P. (2020). Increased baseline RASGRP1 signals Enhance Stem Cell Fitness during Native Hematopoiesis. Oncogene, 39(45), 6920-6934.
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2

Conditional Deletion of Dnmt3a/3b in HSCs

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All animal procedures were IACUC-approved and conducted in accordance the Baylor College of Medicine and Washington University in St. Louis institutional guidelines. All mice were C57Bl/6 background distinguished by CD45.1 or CD45.2 alleles. Dnmt3afl/fl and Dnmt3bfl/fl mice were obtained from the Beaudet lab at Baylor College of Medicine (with the consent of En Li) and crossed to Mx1-cre mice. For bone marrow transplantation, recipient C57Bl/6 CD45.1 mice were transplanted by retroorbital injection following a split dose of 10.5 Gy of lethal irradiation. 250 donor HSCs (CD45.2) were competed against 2.5 × 105 WBM cells with the opposite CD45 allele (matched to the recipient). In competitive transplantation experiments, deletion of floxed alleles in donor HSCs was mediated 5–6 weeks post-transplantation in primary recipients. Conditional deletion was mediated by six intraperitoneal injections (300µg / mouse) of polyinosinic-polycytidylic acid (pIpC; Sigma, St Louis, MO) in PBS every other. For serial HSC transplantation, WBM from transplanted recipients was isolated 18-weeks post-transplant and donor HSCs were re-purified using CD45.2+SPKLS gating. 250 of these re-purified donor HSCs were competed against 2.5 × 105 fresh CD45.1 WBM. PCR screening of Dnmt3a and Dnmt3b floxed allele deletion was performed as previously described (Tadokoro et al., 2007 (link)).
Challen G.A., Sun D., Mayle A., Jeong M., Luo M., Rodriguez B., Mallaney C., Celik H., Yang L., Xia Z., Cullen S., Berg J., Zheng Y., Darlington G.J., Li W, & Goodell M.A. (2014). Dnmt3a and Dnmt3b have Overlapping and Distinct Functions in Hematopoietic Stem Cells. Cell stem cell, 15(3), 350-364.
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3

Conditional Knockout of Dnmt3a and Dnmt3b in Mice

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Animal procedures were approved by the Institutional Animal Care and Use Committee (IACUC) and conducted in accordance with Washington University School of Medicine institutional guidelines. Mice were C57Bl/6 background, Mx1-Cre:Dnmt3afl/fl and Mx1-Cre:Dnmt3afl/fl;Dnmt3bfl/fl mice have been described (16 ). Recombination of floxed alleles was mediated by six intraperitoneal injections (300μg/mouse) of polyinosinic-polycytidylic acid (pIpC; Sigma, St. Louis, MO, USA) in PBS every other day. The following strains were obtained from the Jackson Laboratory (Bar Harbor, ME, USA) - Lck-CRE (003802), CD8a-CRE (008766), Rosa26NICD (008159), Rosa26EYFP (006148) and Nr4a1KO (006187).
Kramer A.C., Kothari A., Wilson W.C., Celik H., Nikitas J., Mallaney C., Ostrander E.L., Eultgen E., Martens A., Valentine M.C., Young A.L., Druley T.E., Figueroa M.E., Zhang B, & Challen G.A. (2017). Dnmt3a Regulates T-cell Development and Suppresses T-ALL Transformation. Leukemia, 31(11), 2479-2490.
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4

Conditional Deletion of hnRNP L in Mice

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hnRNP L floxed mice were described previously26 (link). MxCre, VavCre, H2kBcl2 and Trp53−/− mice were obtained from Jackson laboratories or from a colony maintained at the IRCM. The animal ethics committee of the IRCM approved all animal experiments. MxCre+hnRNP Lfl/fl or hnRNP Lfl/fl mice were injected intraperitoneally with 500 μg of polyinosinic-polycytidylic acid (pIpC; Sigma-Aldrich) every other day for a total of 5 times and were analyzed 14 days after the first injection. All mice were aged between 8–16 weeks for BM analysis.
Gaudreau M.C., Grapton D., Helness A., Vadnais C., Fraszczak J., Shooshtarizadeh P., Wilhelm B., Robert F., Heyd F, & Möröy T. (2016). Heterogeneous Nuclear Ribonucleoprotein L is required for the survival and functional integrity of murine hematopoietic stem cells. Scientific Reports, 6, 27379.
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5

Conditional Yy1 Knockout Mice

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Yy1f/f mice (Affar et al., 2006 (link); Liu et al., 2007 (link)) in which the Yy1 promoter region and exon 1 are flanked by loxP sites, were crossed to Mx1-Cre mice to generate heterozygous Yy1f/+Mx1-cre mice. Yy1f/+Mx1-cre mice were then subsequently bred with Yy1f/f mice to generate homozygous Yy1f/fMx1-cre mice. To induce the expression of Cre recombinase, 6- to 8-week-old mice were injected with 200 mg of polyinosinic-polycytidylic acid (pI-pC; Sigma-Aldrich) every other day for 5 doses. Approximately equal portions of mice of both genders were used, and aggregated data are presented because gender specific differences were not found. All experiments described in this manuscript were performed 7–10 days after the last injection of pI-pC unless stated otherwise. All experiments involving mice were approved by the Institutional Laboratory Animal Care and Use Committee of the University of Wisconsin-Madison and conform to the appropriate regulatory standards.
Lu Z., Hong C.C., Kong G., Assumpção A.L., Ong I.M., Bresnick E.H., Zhang J, & Pan X. (2018). Polycomb Group Protein YY1 Is an Essential Regulator of Hematopoietic Stem Cell Quiescence. Cell reports, 22(6), 1545-1559.
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6

IGF-I Knockout Mice Generation

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The LI-IGF-I−/− mice on a C57BL/6 background were generated as described previously [1 (link), 2 (link)]. Mice homozygous for LoxP [26 (link)] and hemizygous for Mx-Cre [27 (link)] received three ip injections of polyinosinic-polycytidylic acid (PiPc; 6.25 µg/g body weight; Sigma-Aldrich Corp., Stockholm, Sweden) at 3 months of age to induce expression of the Cre protein, thereby inactivating the IGF-I gene in the liver [27 (link)]. PiPc-treated female and male littermates, homozygous for LoxP but lacking Mx-Cre, were used as controls as previously described [2 (link), 3 (link), 26 (link)]. Seven days after the PiPc injections as well as at the end of the study, serum was obtained and assayed for IGF-I by a double-antibody IGF-binding protein-blocked RIA (Mediagnost, Tübingen, Germany). The mice were housed in a standard animal facility under controlled temperature (22 °C) and photoperiod (12 h of light, 12 h of dark) with free access to water and food pellets (B&K Universal AB, Sollentuna, Sweden). Animal care was in accordance with institutional guidelines. The ethical committee at the University of Gothenburg approved this study.
Svensson J., Sjögren K., Levin M., Borén J., Tivesten Å, & Ohlsson C. (2015). Increased diet-induced fatty streak formation in female mice with deficiency of liver-derived insulin-like growth factor-I. Endocrine, 52, 550-560.
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7

Conditional hRasGRP1 Expression Model

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The expression construct was generated by cloning the various components into targeting ROSA vector (ROSA-HR). Vector was obtained by introducing first a flanked Neo/STOP cassette and then the hRasGRP1 cDNA cassette, IRES-EGFP, and human polA sequences into a Rosa26 targeting vector that already contain a ubiquitous CAG promoter cassette (Figure. 1A). This construct was transfected into ES cells by electroporation in optimized condition (5×106 cells in presence of 40 μg linearized plasmid, 260V, 500 uF). Positive selection started 48 h after electroporation by adding 200 μg/ml of G418 in 96-well plates. Positive clones were further screened by southern blotting. Furthermore, the hybridization with internal probe did not indicate that any of the ES cells screened contained an additional randomly integrated copy of the targeting plasmid. At the age of 4 weeks mice received a single intraperitoneal injection of polyinosinic-polycytidylic acid (pIpC) (1μg/250μl/mouse) (Sigma, San Luis, MO, US) to induce the expression of hRASGRP1 and GFP from the Rosa26 locus.
Karra L., Romero-Moya D., Ksionda O., Krush M., Gu Z., Mues M., Depeille P., Mullighan C, & Roose J.P. (2020). Increased baseline RASGRP1 signals Enhance Stem Cell Fitness during Native Hematopoiesis. Oncogene, 39(45), 6920-6934.
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8

Conditional Deletion of miR-21 in Mice

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Normal, wild-type (WT) C57BL/6J mice were purchased from the Institute of Zoology (Chinese Academy of Sciences, Beijing, China). miR-21flox/+ (miR-21fl/+) mice and Mx1-Cre mice were obtained from Shanghai Model Organisms Center (China). miR- 21fl/fl;Mx1-Cre mice were generated by crossing miR-21fl/fl mice with Mx1-Cre mice. Unless otherwise stated, miR-21 deletion was induced by intraperitoneally injecting 4- to 6-week old miR- 21fl/fl;Mx1-Cre+mice with 250 g of polyinosinic:polycytidylic acid (pIpC) (Sigma, St. Louis, MO, USA) every other day for a total of seven doses. Four weeks after pIpC treatment, these mice were used in subsequent experiments. Identically treated miR- 21fl/fl;Mx1-Cre- littermates served as controls. Congenic C57Bl/6 SJL CD45.1+ mice were kindly provided by Prof. Jinyong Wang (Guangzhou Institutes of Biomedicine and Health, Chinese Academy of Science, Guangzhou, China). All animal experiments were approved by the Animal Care Committee of The Third Military Medical University (Chongqing, China).
Hu M., Lu Y., Zeng H., Zhang Z., Chen S., Qi Y., Xu Y., Chen F., Tang Y., Chen M., Du C., Shen M., Wang F., Su Y., Wang S, & Wang J. (2020). MicroRNA-21 maintains hematopoietic stem cell homeostasis through sustaining the nuclear factor-B signaling pathway in mice. Haematologica, 106(2), 412-423.
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9

Conditional Deletion of Fbxo22 in Mice

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Fbxo22fl/fl mice and C57BL/6 Bach1 knockout mice were established at the Cyagen Biosciences Inc. Fbxo22fl/fl mice were further crossed with Myxovirus resistance protein 1 (Mx1)-Cre or stem cell leukemia stem-cell enhancer (Scl)-Cre transgenic mice to achieve specifically inducible deletion of Fbxo22 in hematopoietic cells. All these strains were maintained on a C57BL/6 background. To induce Mx1-Cre recombinase, polyinosinic–polycytidylic acid (pIpC, Sigma, 1 mg/ml in distilled water) was administered at 10 µg/g body weight every other day by intraperitoneal injection for 7 times; to induce Scl-Cre recombinase, tamoxifen (Sigma, 10 mg/ml in corn oil) was administered at 50 µg/g body weight daily by intraperitoneal injection for 21 days. All the animal experiments were approved by our institution and conducted according to the Guideline for Animal Care at Shanghai Jiao Tong University School of Medicine.
Zhu X.N., Wei Y.S., Yang Q., Liu H.R., Zhi Z., Zhu D., Xia L., Hong D.L., Yu Y, & Chen G.Q. (2023). FBXO22 promotes leukemogenesis by targeting BACH1 in MLL-rearranged acute myeloid leukemia. Journal of Hematology & Oncology, 16, 9.
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10

Conditional Yy1 Knockout Mice

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Yy1f/f mice (Affar et al., 2006 (link); Liu et al., 2007 (link)) in which the Yy1 promoter region and exon 1 are flanked by loxP sites, were crossed to Mx1-Cre mice to generate heterozygous Yy1f/+Mx1-cre mice. Yy1f/+Mx1-cre mice were then subsequently bred with Yy1f/f mice to generate homozygous Yy1f/fMx1-cre mice. To induce the expression of Cre recombinase, 6- to 8-week-old mice were injected with 200 mg of polyinosinic-polycytidylic acid (pI-pC; Sigma-Aldrich) every other day for 5 doses. Approximately equal portions of mice of both genders were used, and aggregated data are presented because gender specific differences were not found. All experiments described in this manuscript were performed 7–10 days after the last injection of pI-pC unless stated otherwise. All experiments involving mice were approved by the Institutional Laboratory Animal Care and Use Committee of the University of Wisconsin-Madison and conform to the appropriate regulatory standards.
Lu Z., Hong C.C., Kong G., Assumpção A.L., Ong I.M., Bresnick E.H., Zhang J, & Pan X. (2018). Polycomb Group Protein YY1 Is an Essential Regulator of Hematopoietic Stem Cell Quiescence. Cell reports, 22(6), 1545-1559.
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