Myd88

Manufactured by Jackson ImmunoResearch

Sourced in Montenegro, United States

Myd88−/− is a laboratory-produced mouse strain that carries a genetic modification resulting in the absence of the Myd88 protein. The Myd88 protein plays a crucial role in immune signaling pathways. This mouse strain is a valuable tool for researchers investigating the function of the Myd88 protein and its impact on the immune system.

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MyD88−/− mice (22 citations) B6.129P2(SJL)-MyD88tm1.1Defr/J (MyD88−/−, (7 citations) MyD88 KO mice (6 citations) MyD88 knockout mice (5 citations) MyD88 knockout (KO) mice (4 citations) MyD88-floxed mice (3 citations) Floxed’ MyD88 (3 citations) B6.129P2(SJL)-MyD88/J mice (2 citations) Myd88−/−24 (2 citations) MyD88-deficient (2 citations)

47 protocols using myd88

Dissecting the Roles of Immune Receptors in MCMV Infection

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All in vivo experiments were performed according to UK Home Office guidelines and were performed at Cardiff University (Reference: P7867DADD) in specific pathogen-free/SPF conditions at ambient temperature and humidity. Nogo-A/B^−/− mice^{73 (link)}, and IFITM3-deficient (Ifitm3^−/−) and wt control mice have been described previously^{1 (link)} and were crossed with Myd88^−/− (Jackson Laboratory) or for some experiments Nogo-A/B^−/− mice to generate Ifitm3^−/−/Myd88^−/− and Ifitm3^−/−/Nogo-A/B^−/− mice. Novel strains are available upon request. Age- and sex-matched mice between 7 and 12 weeks of age were used in the experiments. Tlr3^−/−^{74 (link)}, Tlr7^−/−^{75 (link)} and Tlr9^−/−^{76 (link)} mice were a kind gift from Caetano Reis e Sousa (Crick Institute, London). Relevant wild type control mice were all bred inhouse. MCMV (pSM3fr-MCK-2fl BACmid) was grown and titred using 3T3 cells (ATCC, CRL-1658) with a carboxycellulose overlay. Mice were infected via intraperitoneal (i.p.) injection with between 5 × 10⁵ to 2 × 10⁶ PFU MCMV. For Anakinra treatment, mice were injected i.p. with Anakinra (KINERET: Cardiff & Vale NHS Pharmacy) (25 mg/kg) or PBS control on day 0 p.i. For infectious virus quantification form harvested tissue, viral load was determined via plaque assay as previously described^{77 (link)}. All mice were euthanized using carbon dioxide.

Clement M., Forbester J.L., Marsden M., Sabberwal P., Sommerville M.S., Wellington D., Dimonte S., Clare S., Harcourt K., Yin Z., Nobre L., Antrobus R., Jin B., Chen M., Makvandi-Nejad S., Lindborg J.A., Strittmatter S.M., Weekes M.P., Stanton R.J., Dong T, & Humphreys I.R. (2022). IFITM3 restricts virus-induced inflammatory cytokine production by limiting Nogo-B mediated TLR responses. Nature Communications, 13, 5294.

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Immune Signaling Pathways in Murine Studies

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Wild-type (WT) C57BL/6, TLR2^−/−, MyD88^−/−, μMT, and STING^gt/gt mice were purchased from The Jackson Laboratory. TLR9^−/− mice had been kindly provided by Dr. Daniel Muruve (University of Calgary) and were bred in house at the University of Florida [18 (link)]. IFNαR^−/− mice were housed at Baylor College of Medicine as previously described [23 (link)]. All knockout mice were on a C57BL/6 background. Animals were housed under specific pathogen free conditions at the University of Florida or Baylor College of Medicine and treated under Institutional Animal Care and Use Committee approved protocols. All animals were male and 6-8 weeks old at the onset of the experiments; all cohorts contained at least 4 mice per group.
AAV vectors were administered intramuscularly or intravenously as previously described [18 (link),20 (link)]. Plasma samples were collected by retro-orbital bleed into heparinized capillary tubes. TLR9i (ODN 2088, Invivogen) was delivered at 100 μg/mouse mixed with the vector formulation as previously described [18 (link)]. MyD88 inhibitor (MyD88i) and control peptide (MyD88c) (IMG-2005, Imgenex) were delivered at 25 μg/mouse i.p. in 100 μL PBS as previously described [24 (link)].

Rogers G.L., Suzuki M., Zolotukhin I., Markusic D.M., Morel L.M., Lee B., Ertl H.C, & Herzog R.W. (2015). Unique Roles of TLR9- and MyD88-Dependent and -Independent Pathways in Adaptive Immune Responses to AAV-Mediated Gene Transfer. Journal of Innate Immunity, 7(3), 302-314.

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Murine Pancreatic Cancer Cell Lines

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6–8 week old female C57BL/6 (000664), IL12p35^−/− (002692), IFNγ^−/− (002287), MyD88^−/− (009088) and CD8a^−/− (002665) were purchased from Jackson Laboratory. All animal work was performed at the Dartmouth Hitchcock Medical Center animal facility with Dartmouth IACUC approval. The murine pancreatic adenocarcinoma Pan02 cell line, also known as Panc02 (26 (link)), was acquired from the Division of Cancer Treatment Tumor Repository (NCI). Pan02 cells were maintained in high glucose Roswell Park Memorial Institute (RPMI) 1640 media. ID8-GFP cells (27 (link)) were maintained in high glucose Dulbecco's Modified Eagle Medium (DMEM). Human foreskin fibroblasts (HFF) (28 (link)) cultures were maintained in Eagle's Minimum Essential Medium (EMEM). All cell culture media was supplemented with 10% FBS, L-glutamine, and penicillin/streptomycin.

Sanders K.L., Fox B.A, & Bzik D.J. (2015). Attenuated Toxoplasma gondii stimulates immunity to pancreatic cancer by manipulation of myeloid cell populations. Cancer immunology research, 3(8), 891-901.

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Genetically Modified Mice for Immunological Research

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C57BL/6 mice (strain #664), B6129PF2/J (strain #100903), Trpa1^–/– (strain #6401), Tlr7^–/– (strain #8380), Trpv1^–/– (strain #3770), Il-12p40^–/– (strain #2693), Tcrd^–/– (strain #2120), Myd88^–/– (strain #9088), Nlrp3^–/– (strain # 21302), CD11c-DTR/GFP (strain #4509), Scid (strain #1913), and Foxn1^–/–J:NU (strain #7850) were purchased from The Jackson Laboratory. All mice were group housed in the animal facility of University of Pennsylvania on a 12-hour light/12-hour dark cycle with ad libitum access to water and normal chow. Mice were verified by genotyping. The CD11c-DTR/GFP transgenic line has the CD11c promoter directing expression of a diphtheria toxin receptor - enhanced green fluorescent protein (DTR/EGFP) fusion protein to dendritic cell populations. To deplete dendritic cell populations, we administered diphtheria toxin intraperitoneal (100ng per mouse) 3 times per week. Mice used in this study were male and female age-matched and background-matched littermates that were randomly assigned to experimental groups. No sex-dependent differences were observed.

Wei J.J., Kim H.S., Spencer C.A., Brennan-Crispi D., Zheng Y., Johnson N.M., Rosenbach M., Miller C., Leung D.H., Cotsarelis G, & Leung T.H. (2020). Activation of TRPA1 Nociceptor Promotes Systemic Adult Mammalian Skin Regeneration. Science immunology, 5(50), eaba5683.

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Mouse Model Comparison for Immune Studies

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BALB/c (catalog no. 000651), CBA/J (catalog no. 000656), C57BL/6 (catalog no. 000664), Card9^−/− (catalog no. 028652), and MyD88^−/− (catalog no. 009088) mice were obtained from Jackson Laboratory (Bar Harbor, ME). BALB/c and C57BL/6 mice obtained from Jackson Laboratory are also known as BALB/cJ and C57BL/6 mice, respectively. All mice were 6 to 8 weeks old at the time of inoculation. All animal protocols were reviewed and approved by the Animal Studies Committee of the Washington University School of Medicine and conducted according to National Institutes of Health guidelines for housing and care of laboratory animals.

Hole C.R., Lam W.C., Upadhya R, & Lodge J.K. (2020). Cryptococcus neoformans Chitin Synthase 3 Plays a Critical Role in Dampening Host Inflammatory Responses. mBio, 11(1), e03373-19.

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Murine Plasmodium Infection Model

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Female mice of C57BL/6 (WT), Aim2^−/−, Casp1^−/−, Myd88^−/−, Nlrp3^−/−, Il1r1^−/− and Zbtb46-DTR mice were purchased from The Jackson Laboratory. Traf3^flox/flox mice were kindly gifted from Dr. Shao-Cong Sun (University of Texas, MD Anderson Cancer Center) and cross with CD11c-cre (The Jackson Laboratory) to generate Traf3^f/f CD11c-cre mice, Irf3^−/−:Irf7^−/− mice were from Dr. Kate Fitzgerald (University of Massachusetts Medical School) and Dr. Tadatsugo Taniguchi (The University of Tokyo), and crossed with C57BL/6 mice to get Irf3^−/− mice. For plasmodium infection, 0.5 × 10⁶ iRBCs (otherwise, indicated specifically in the figure legend) suspended in 200 µl PBS from the donor mice were intraperitoneally injected into experimental mice. All mouse-related procedures were performed according to experimental protocols approved by the Animal Care and Welfare Committee at Houston Methodist Research Institute and in accordance with NIH-approved animal study protocol LMVR-11E.

Yu X., Du Y., Cai C., Cai B., Zhu M., Xing C., Tan P., Lin M., Wu J., Li J., Wang M., Wang H.Y., Su X.Z, & Wang R.F. (2018). Inflammasome activation negatively regulates MyD88-IRF7 type I IFN signaling and anti-malaria immunity. Nature Communications, 9, 4964.

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Generating Genetically Engineered Mice

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Wild type (WT) C57BL/6J, TLR2^−/−, MyD88^−/−, and ApoE^−/− mice were purchased from Jackson Laboratories (Bar Harbor, ME). CD36^−/− mice were generated as described.^{27 (link)}TLR6^−/− mice were obtained from Oriental Yeast Co. LTD (Tokyo, Japan). All strains were of at least 99% C57BL/6J background. Src^−/− and Lyn^−/− mice were generated as reported earlier.^{9 (link)} We generated ApoE^−/−/CD36^−/− by crossing ApoE^−/− with CD36^−/− mice as described^{10 (link)}, and ApoE^−/−/TLR2^−/− and ApoE^−/−/TLR6^−/− mice by crossing ApoE^−/− with TLR2^−/− or TLR6^−/− mice respectively. We further crossed ApoE^−/−/CD36^−/− mice to ApoE^−/−/TLR2^−/− or ApoE^−/−/TLR6^−/− mice to generate ApoE^−/−/CD36^−/−/TLR2^−/− or ApoE^−/−/CD36^−/−/TLR6^−/− triple knockout mice. Mice were fed a Western diet (TD88137, Harlan Teklad) for 8–10 weeks. Animals were housed in ventilated cages with ad libitum access to food and water, on a 14:10 light dark cycle. Prior to study, the Institutional Animal Care and Use Committee of the Cleveland Clinic approved all animal procedures. Mice were used between 8 and 14 weeks of age.

Biswas S., Zimman A., Gao D., Byzova T.V, & Podrez E.A. (2017). TLR2 Plays a Key Role in Platelet Hyperreactivity and Accelerated Thrombosis Associated with Hyperlipidemia. Circulation research, 121(8), 951-962.

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Genetically Modified Mice for Immunological Studies

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WT C57BL/6J, MyD88^−/−, MyD88^fl, Cγ1-cre mice (all on the C57BL/6J background), and BALB/c mice, were obtained from The Jackson Laboratory (Bar Harbor, ME). The alpha-galactosyl transferase^−/− mice (GT KO) were a generous gift from Dr. Uri Galili (Rush University, Chicago, IL). All of the mice were maintained at the University of Virginia, under specific pathogen-free conditions and used according to the regulations and standard guidelines of the Institutional Animal Care and Use Committee. All experiments were conducted with 8-10-week old female mice unless otherwise noted.

Chandrasekhar J.L., Cox K.M., Loo W.M., Qiao H., Tung K.S, & Erickson L.D. (2019). Cutaneous Exposure to Clinically Relevant Lone Star Ticks Promote IgE Production and Hypersensitivity Through CD4+ T Cell- and MyD88-Dependent Pathways in Mice. Journal of immunology (Baltimore, Md. : 1950), 203(4), 813-824.

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Generating Transgenic Mouse Models

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Six- to eight-week old C57BL/6J mice were purchased from Harlan. Myd88^−/−, Trif^−/−, Camp^−/−, 2C CD8⁺ T cell receptor (TCR)-Tg, Cd11c^Cre+-Tg mice were purchased from The Jackson Laboratory. Ifnar1^flox/flox mice were kindly provided by Dr. Ulrich Kalinke of the Institute for Experimental Infection Research, Hanover, Germany. Tmem173^−/− mice were kindly provided by Dr. Glen N. Barber of University of Miami School of Medicine, Miami. Irf3^−/− mice were kindly provided by T. Taniguchi of University of Tokyo, Tokyo, Japan. All the mice were maintained under specific pathogen free conditions and used in accordance to the animal experimental guidelines set by the Institute of Animal Care and Use Committee. This study has been approved by the Institutional Animal Care and Use Committee of the University of Chicago.

Deng L., Liang H., Xu M., Yang X., Burnette B., Arina A., Li X.D., Mauceri H., Beckett M., Darga T., Huang X., Gajewski T.F., Chen Z.J., Fu Y.X, & Weichselbaum R.R. (2014). STING-dependent Cytosolic DNA Sensing Promotes Radiation-induced Type I interferon-dependent Antitumor Immunity in Immunogenic Tumors. Immunity, 41(5), 843-852.

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Gnotobiotic Mouse Model for Microbiome Studies

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Male Swiss Webster (SW)/Germ free (GF) mice (6–8 weeks old) were obtained from Taconic Farms (Germantown, NY). Experiments with gnotobiotic mice were carried out in sterile micro-isolators. Water and a commercial autoclavable diet were sterilized by steam and administered ad libitum to all the animals. To confirm sterility of germ free mice, fecal samples were cultured using a thioglycolate test every two days. Male C57BL/6J, and MyD88^−/− (6–8 weeks) were purchased from Jackson Laboratory (Bar Harbor, ME). The mice were housed under specific pathogen-free conditions in standard cages and fed standard laboratory chow and water. All animal procedures were reviewed and approved by the Institutional Animal Care and Use Committee of Louisiana State University Health Sciences Center-Shreveport. All animal experiments were performed according to the criteria outlined by the National Institutes of Health.

Souza D.G., Senchenkova E.Y., Russell J, & Granger D.N. (2015). MyD88 mediates the protective effects of probiotics against the arteriolar thrombosis and leukocyte recruitment associated with experimental colitis. Inflammatory bowel diseases, 21(4), 888-900.

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