The interaction between TLR9 and downstream molecule myeloid differentiation 88 (MyD88) was examined by immunoprecipitation. After HEK-TLR9 cells were stimulated with ODN2006 for 3 hours with or without isoflurane (2%), sevoflurane (2%), or propofol (100 μM), they were subjected to lysis with RIPA buffer containing protease inhibitor cocktail Complete. Following centrifugation, supernatants were subjected to immunoprecipitation using rabbit anti-human TLR9 antibody (Cell Signaling Technology; Danvers, MA, USA) and Dynabeads Protein A immunoprecipitation kit (Thermofisher Scientific; Waltham, MA, USA). Bead pellets were subjected to western blot analysis and probed with mouse anti-human MyD88 antibody (Santa Cruz Biotechnology Inc; Dallas, TX, USA) followed by antimouse IgG-horseradish peroxidase (HRP) antibody (Cell Signaling). Immunoreactive proteins were visualized by enhanced chemiluminescence.
Mouse anti human myd88 antibody
Manufactured by Santa Cruz Biotechnology
Sourced in United States
The Mouse anti-human MyD88 antibody is a laboratory reagent designed for use in research applications. It specifically binds to the human MyD88 protein, which is a key adaptor molecule involved in Toll-like receptor and interleukin-1 receptor signaling pathways. This antibody can be utilized in various immunoassay techniques to detect and study the MyD88 protein.
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Lab products found in correlation
3 protocols using mouse anti human myd88 antibody
1
TLR9-MyD88 Interaction Modulation
2
Visualizing MyD88 Localization in Cells
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PaKiT03 cells were seeded onto glass coverslips in 24-well plates at 105 cells per well one day before transfection. They were transfected with 200 ng each of GFP-IRF7 and FLAG-tagged MyD88 (human or bat) using Lipofectamine 2000 (Life Technologies). At 16 h post-transfection, cells were fixed in 4% paraformaldehyde in PBSA at room temperature for 40 minutes. After removal of fixative, cells were washed three times with PBSA, followed by treatment with 0.1% Triton X-100 for 10 minutes and blocked with 0.5% BSA in PBSA for 30 minutes. Mouse anti-human MyD88 antibody (Santa Cruz, CAT. sc11356) was diluted 1∶1000 in 0.5% BSA and applied to cells and incubated at 37°C for 1 h. Cells were then incubated with Alexa 488 conjugated goat anti-mouse secondary antibody at 37°C for 1 h and washed three times in PBSA. Nuclei were labelled with DAPI and coverslips were mounted on glass slides for analysis. All slides were examined under a Leica confocal microscope (Leica, Germany).
3
TLR7-MyD88 Interaction Examined by Immunoprecipitation
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