Detects human TLR4 in direct ELISAs and Western blots. In direct ELISAs, no cross-reactivity with recombinant human (rh) TLR1, 2, 3, 5, 7, 8, 10, recombinant mouse (rm) TLR4/MD-2 complex, rmTLR6, rhIL-1RI, rhIL-1RII, rhIL-1RAcP, rhST2, rhIL-18R, rhIL-1Rrp2, rhIL-18AcPLR, rhSIGIRR, rhTIGIRR, rhMD-1, rhMD-2, or rmMD-2 is observed.
Monoclonal Mouse IgG1 Clone # 610029
Protein A or G purified from hybridoma culture supernatant
Mouse myeloma cell line NS0-derived recombinant human TLR4 Glu24-Lys631 Accession # O00206
Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. *Small pack size (SP) is supplied as a 0.2 µm filtered solution in PBS.
Detection of Human TLR4 by Western Blot.
Western blot shows lysates of HeLa human cervical epithelial carcinoma cell line. PVDF Membrane was probed with 2 µg/mL of Mouse Anti-Human TLR4 Monoclonal Antibody (Catalog # MAB14782) followed by HRP-conjugated Anti-Mouse IgG Secondary Antibody (Catalog # HAF007). A specific band was detected for TLR4 at approximately 120 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.
Preparation and Storage
Reconstitute at 0.5 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.
TLR4 is a 100 kDa type I transmembrane glycoprotein that belongs to the mammalian Toll-Like Receptor family of pathogen pattern recognition molecules. MD-2, also known as ESOP-1, is a 25 kDa secreted protein that is required for TLR4-mediated responses to bacterial lipopolysaccharide (LPS) (1‑3). The human TLR4 cDNA encodes an 839 amino acid (aa) precursor that contains a 23 aa signal sequence, a 608 aa extracellular domain (ECD), a 21 aa transmembrane segment, and a 187 aa cytoplasmic domain. TLR4 contains 21 leucine rich repeats in its ECD and one cytoplasmic Toll/IL-1 receptor (TIR) domain (4). The ECD of human TLR4 shares approximately 25% aa sequence identity with other TLRs and 60%‑74% aa sequence identity with bovine, equine, feline, mouse, rat, and porcine TLR4. The human MD-2 cDNA encodes a 160 aa precursor with an 18 aa signal sequence (5). Human MD-2 shares 20% aa sequence identity with MD-1 and 62%‑64% aa sequence identity with bovine, mouse, and rat MD-2. MD-2 associates with TLR4 on monocytes, macrophages, dendritic cells, and B cells (5‑7). MD-2 expression is required for cell surface localization of TLR4 and for optimal LPS-induced TLR4 signaling (7, 8). MD-2 also forms soluble disulfide-linked homo-oligomers which can interact with TLR4 (6). Through a domain separate from its TLR4-binding domain, MD-2 extracts LPS from circulating CD14-LPS complexes and carries the LPS into a ternary complex with TLR4 (9‑11). The interaction of MD-2/LPS with TLR4 induces receptor oligomerization and the triggering of an inflammatory response (12). Increased levels of plasma MD-2 in septic shock patients sensitizes MD-2 non-expressing epithelial cells to LPS and promotes widespread tissue inflammation (13).
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