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Anti mouse myd88

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Anti-mouse MyD88 is a laboratory reagent used in research applications. It is an antibody that specifically binds to the MyD88 protein in mouse samples. MyD88 is an adaptor protein involved in the innate immune response. The antibody can be used to detect and study the expression and localization of MyD88 in mouse-derived cells and tissues.

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Lab products found in correlation

Lysis buffer, by Cell Signaling Technology (2 mentions) Anti mouse tlr7, by Thermo Fisher Scientific (2 mentions) Anti mouse igg isotype control, by Santa Cruz Biotechnology (2 mentions) Anti glyceraldehyde 3 phosphate dehydrogenase gapdh, by Cell Signaling Technology (1 mentions) Horseradish peroxidase hrp, by Agilent Technologies (1 mentions) Anti bcl 2 associated x protein bax, by Santa Cruz Biotechnology (1 mentions) Fluorescein isothiocyanate conjugated anti rabbit antibody, by Abcam (1 mentions) Caco 2 cells, by European Collection of Cell Cultures (1 mentions) Mouse anti nfkb, by Santa Cruz Biotechnology (1 mentions) V3 western workflow, by Bio-Rad (1 mentions) Peroxidase conjugated anti mouse secondary antibody, by Fortis Life Sciences (1 mentions) Alliance 2.7 system, by Uvitec (1 mentions)

4 protocols using anti mouse myd88

1

TLR7-MyD88 Co-immunoprecipitation in BMDCs

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Co-immunoprecipitation experiments were performed using BMDCs seeded in 6-well plates at a final concentration of 1×106 cells/mL in RPMI-complete media. Cells were incubated with 10 μg/mL Gardiquimod for 15 minutes at 37C with 5% CO2. Cells were harvested and lysed in 1× lysis buffer (Cell Signaling). Lysates were incubated with anti-mouse TLR7 (Fisher Scientific) or anti-mouse IgG isotype control (Santa Cruz Biotech) for 24 h at 4C. Supernatants were incubated with Protein G agarose beads overnight at 4C. Precipitated proteins are eluted, separated by SDS-PAGE and immunoblotted with anti-mouse MyD88 (Santa Cruz Biotech).
Ramirez-Ortiz Z.G., Prasad A., Griffith J.W., Pendergraft WF I.I.I., Cowley G.S., Root D.E., Tai M., Luster A.D., Khoury J.E., Hacohen N, & Means T.K. (2015). TREML4 amplifies TLR7-mediated signaling during antiviral responses and autoimmunity. Nature immunology, 16(5), 495-504.
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2

TLR7-MyD88 Co-immunoprecipitation in BMDCs

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Co-immunoprecipitation experiments were performed using BMDCs seeded in 6-well plates at a final concentration of 1×106 cells/mL in RPMI-complete media. Cells were incubated with 10 μg/mL Gardiquimod for 15 minutes at 37C with 5% CO2. Cells were harvested and lysed in 1× lysis buffer (Cell Signaling). Lysates were incubated with anti-mouse TLR7 (Fisher Scientific) or anti-mouse IgG isotype control (Santa Cruz Biotech) for 24 h at 4C. Supernatants were incubated with Protein G agarose beads overnight at 4C. Precipitated proteins are eluted, separated by SDS-PAGE and immunoblotted with anti-mouse MyD88 (Santa Cruz Biotech).
Ramirez-Ortiz Z.G., Prasad A., Griffith J.W., Pendergraft WF I.I.I., Cowley G.S., Root D.E., Tai M., Luster A.D., Khoury J.E., Hacohen N, & Means T.K. (2015). TREML4 amplifies TLR7-mediated signaling during antiviral responses and autoimmunity. Nature immunology, 16(5), 495-504.
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3

Caco-2 Cell Culture and Western Blot Analysis

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Caco-2 cells were purchased from European Collection of Cell Cultures (ECACC, Public Health England Porton Down, Salisbury, UK). Cell medium, chemicals and reagents used for cell culture, and TcdA were purchased from Sigma–Aldrich (St. Louis, MO, USA), unless otherwise stated. Instruments, reagents, and materials used for western blot analysis were obtained from Bio-Rad Laboratories (Milan, Italy). Rabbit anti-zona occludens-1 (ZO-1), anti-occludin and anti-glyceraldehyde-3-phosphate dehydrogenase (GAPDH) antibodies were procured from Cell Signaling Technology (Danvers, MA, USA). Rabbit anti-toll-like receptor 4 (TLR4), mouse anti-ZO-1, anti-Bcl-2-associated X protein (Bax), mouse anti-MyD88, rabbit anti-transforming growth factor-β-activated kinase-1 (pTAK1), and mouse anti-TAK1 antibody were purchased from Santa Cruz Biotechnology (Santa Cruz, CA, USA) and horseradish peroxidase (HRP) was obtained from Dako (Milan, Italy). Fluorescein isothiocyanate-conjugated anti-rabbit antibody and Texas red conjugated anti-mouse antibody were purchased from Abcam (Cambridge, UK), and custom oligonucleotides for electrophoretic mobility shift assay (EMSA) analysis were synthesized by TIB Molbiol (Berlin, Germany).
Esposito G., Nobile N., Gigli S., Seguella L., Pesce M., d’Alessandro A., Bruzzese E., Capoccia E., Steardo L., Cuomo R, & Sarnelli G. (2016). Rifaximin Improves Clostridium difficile Toxin A-Induced Toxicity in Caco-2 Cells by the PXR-Dependent TLR4/MyD88/NF-κB Pathway. Frontiers in Pharmacology, 7, 120.
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4

Profiling Inflammatory Mediators in hGFs

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Proteins from untreated and MPs-treated hGFs (for 48 h) were separated using sodium dodecyl-sulfate polyacrylamide gel electrophoresis (SDS-PAGE) followed by Western blot analysis (Bio-Rad V3 Western Workflow™, Milan, Italy). Membranes were saturated for 120 min at room temperature in a blocking buffer (1 × TBS, 5% milk, 0.1% Tween-20) followed by overnight incubation at 4 °C with the following primary antibodies: mouse anti-NFkB (1:500; Santa Cruz Biotechnology), mouse anti-MyD88 (1:500; Santa Cruz Biotechnology) and mouse anti-NLRP3 (3 µg/mL; Novus). Subsequently, membranes were incubated for 60 min at room temperature with peroxidase-conjugated anti-mouse secondary antibody (1:5000; Bethyl Laboratories, Montgomery, AL, USA) [35 (link)]. Enhanced chemiluminescence with the Alliance 2.7 system (Uvitec Ltd., Cambridge, UK) was used to identify and quantify the bands obtained.
Caputi S., Diomede F., Lanuti P., Marconi G.D., Di Carlo P., Sinjari B, & Trubiani O. (2022). Microplastics Affect the Inflammation Pathway in Human Gingival Fibroblasts: A Study in the Adriatic Sea. International Journal of Environmental Research and Public Health, 19(13), 7782.
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