Mouse/Rat MyD88 Antibody

Detection of Mouse/Rat MyD88 by Western Blot. Western blot shows lysates of L6 rat myoblast cell line, A20 mouse B cell lymphoma cell line, and CH-1 mouse B cell lymphoma cell line. PVDF membrane was probed with 1 µg/mL of Goat Anti-Mouse/Rat MyD88 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF3109) followed by HRP-conjugated Anti-Goat IgG Secondary Antibody (Catalog # HAF017). A specific band was detected for MyD88 at approximately 34 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 2.

MyD88 in RAW 264.7 Mouse Cell Line. MyD88 was detected in immersion fixed RAW 264.7 mouse monocyte/macrophage cell line using Goat Anti-Mouse/Rat MyD88 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF3109) at 1.7 µg/mL for 3 hours at room temperature. Cells were stained using the NorthernLights™ 557-conjugated Anti-Goat IgG Secondary Antibody (red; Catalog # NL001) and counterstained with DAPI(blue). Specific staining was localized to cytoplasmic. View our protocol for Fluorescent ICC Staining of Cells on Coverslips.

Detection of MyD88 in Mouse Splenocytes by Flow Cytometry. Mouse splenocytes were stained with Goat Anti-Mouse/Rat MyD88 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF3109, filled histogram) or control antibody (Catalog # AB-108-C, open histogram), followed by Allophycocyanin-conjugated Anti-Goat IgG Secondary Antibody (Catalog # F0108). To facilitate intracellular staining, cells were fixed with paraformaldehyde and permeabilized with saponin. This application has not been tested in rat samples.

Detection of Rat MyD88 by Simple Western^TM. Simple Western lane view shows lysates of Nb2-11 rat lymphoma cell line, L6 rat myoblast cell line, and NR8383 rat alveolar macrophage cell line, loaded at 0.2 mg/mL. A specific band was detected for MyD88 at approximately 40 kDa (as indicated) using 10 µg/mL of Goat Anti-Mouse/Rat MyD88 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF3109) followed by 1:50 dilution of HRP-conjugated Anti-Goat IgG Secondary Antibody (Catalog # HAF109). This experiment was conducted under reducing conditions and using the 12-230 kDa separation system.

Detection of Mouse Mouse/Rat MyD88 Antibody by Western Blot Examination of IRAK1 in myddosome interactions. D, WT iBMDMs expressing IRAK1-mCerulean were left untreated or stimulated with 1 μg/ml LPS for 30 min or 2 h. WCLs were then subjected to GFP IP followed by immunoblotting for MyD88 and GFP or directly subjected to immunoblotting with the same antibodies. MyD88 in WCLs also served as a loading control. The data presented are representative of three independent experiments.Image collected and cropped by CiteAb from the following publication (https://pubmed.ncbi.nlm.nih.gov/30076215), licensed under a CC-BY license. Not internally tested by R&D Systems.

Detection of Mouse Mouse/Rat MyD88 Antibody by Western Blot Examination of IRAK1 in myddosome interactions.A and B, WT iBMDMs expressing IRAK1-mCerulean were left untreated (UT) or stimulated with 1 μg/ml LPS for 2 or 4 h. E, WT iBMDMs expressing IRAK1-mCerulean were pretreated with either DMSO or 20 μm I4 inhb for 30 min. Cells were then left untreated or stimulated with 1 μg/ml LPS for a further 30 min. WCLs were then subjected to GFP IP followed by immunoblotting with MyD88 and GFP or directly subjected to immunoblotting for GFP, P-IRAK4, and MyD88. The data presented are representative of four independent experiments. Image collected and cropped by CiteAb from the following publication (https://pubmed.ncbi.nlm.nih.gov/30076215), licensed under a CC-BY license. Not internally tested by R&D Systems.