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.
Simple Western lane view shows lysates of L6 rat myoblast cell line, loaded at 0.2 mg/mL. A specific band was detected for MyD88 at approximately 39 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 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.
Preparation and Storage
Reconstitute at 0.2 mg/mL in sterile PBS.
Reconstitution Buffer Available
The product is shipped at ambient temperature. Upon receipt, store it immediately at the temperature recommended below. *Small pack size (SP) is shipped with polar packs. Upon receipt, store it immediately at -20 to -70 °C
Stability & Storage
Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
12 months from date of receipt, -20 to -70 °C as supplied.
1 month, 2 to 8 °C under sterile conditions after reconstitution.
6 months, -20 to -70 °C under sterile conditions after reconstitution.
Myeloid Differentiation primary response protein 88 (MyD88) is a 296 amino acid, 34 kDa, ubiquitously expressed, cytoplasmic adaptor protein involved in the signaling of TLR and IL-1 R family members. MyD88 contains an N-terminal death domain and a C-terminal Toll/IL-1 R (TIR) domain. Each domain seems to participate in protein-protein interactions, as the death domain is inactive. Upon Toll receptor ligation, MyD88 is recruited to the receptor and initiates a signaling cascade that results in NK kappa B and JNK activation. The amino acid sequence of mouse MyD88 is 93% identical to that of rat MyD88.
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