Human TLR4 Affinity Purified Polyclonal Ab (25 UG)【科瑞奥博】

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Human TLR4 Affinity Purified Polyclonal Ab (25 UG)

货号： AF1478-SP 基本售价： 1378.1 元规格： -

产品信息

概述
货号	AF1478-SP
别名	ARMD10; CD284 antigen; CD284; hTollhomolog of Drosophila toll; TOLL; toll-like receptor 4
全称	Toll-like Receptor 4
反应种属	Human
应用	Western Blot(1 µg/mL) Flow Cytometry(2.5 µg/10⁶cells) Immunohistochemistry(5-15 µg/mL) Immunocytochemistry(5-15 µg/mL)
目标/特异性	Detects human TLR4 in direct ELISAs and Western blots. In direct ELISAs and Western blots, approximately 30% cross-reactivity with recombinant mouse (rm) TLR4 is observed and less than 5% cross-reactivity with recombinant human (rh) TLR1, rhTLR2, rhTLR3 and rmTLR6 is observed.
使用方法	Western Blot: 1 µg/mL Flow Cytometry: 2.5 µg/10⁶cells Immunohistochemistry: 5-15 µg/mL Immunocytochemistry: 5-15 µg/mL Neutralization: Measured by its ability to neutralize LPS-induced IL‑8 secretion in the HEK293 human embryonic kidney cell line co-transfected with human TLR4 and MD-2. The Neutralization Dose (ND₅₀) is typically 1.5-7.5 µg/mL in the presence of 75 ng/mL Lipopolysacharide (LPS).
来源	Polyclonal Goat IgG
产品组分

性能
供应商	R&D Systems
Entrez Gene IDs	7099 (Human); 21898 (Mouse); 29260 (Rat)
应用文献
R&D Systems personnel manually curate a database that contains references using R&D Systems products. The data collected includes not only links to publications in PubMed, but also provides information about sample types, species, and experimental conditions. The C-terminal disulfide bonds of Helicobacter pylori GroES are critical for IL-8 secretion via the TLR4-dependent pathway in gastric epithelial cells. Authors: Su Y, Yang J, Lee H, Sheu F, Hsu C, Lin S, Chow L J Immunol, 2015;194(8):3997-4007. Species: H. pylori Sample Type: Cell Lysate Application: IP TLR4 Expression Is Associated with Left Ventricular Dysfunction in Patients Undergoing Coronary Artery Bypass Surgery. Authors: Avlas O, Bragg A, Fuks A, Nicholson J, Farkash A, Porat E, Aravot D, Levy-Drummer R, Cohen C, Shainberg A, Arad M, Hochhauser E PLoS ONE, 2015;10(6):e0120175. Species: Human Sample Type: Whole Tissue Application: IHC Frozen HIV-1 Tat Protein Induces Production of Proinflammatory Cytokines by Human Dendritic Cells and Monocytes/Macrophages through Engagement of TLR4-MD2-CD14 Complex and Activation of NF-kappaB Pathway. Authors: Ben Haij N, Planes R, Leghmari K, Serrero M, Delobel P, Izopet J, BenMohamed L, Bahraoui E PLoS ONE, 2015;10(6):e0129425. Species: Human Sample Type: Whole Cells Application: Flow Slit2-Robo4 pathway modulates lipopolysaccharide-induced endothelial inflammation and its expression is dysregulated during endotoxemia. Authors: Zhao H, Anand A, Ganju R J Immunol, 2014;192(1):385-93. Species: Human Sample Type: cells Application: Neut Pathogenic strains of Acanthamoeba are recognized by TLR4 and initiated inflammatory responses in the cornea. Authors: Alizadeh H, Tripathi T, Abdi M, Smith A PLoS ONE, 2014;9(3):e92375. Species: Human Sample Type: Whole Cells Application: Neut Fetal fibronectin signaling induces matrix metalloproteases and cyclooxygenase-2 (COX-2) in amnion cells and preterm birth in mice. Authors: Mogami H, Kishore A, Shi H, Keller P, Akgul Y, Word R J Biol Chem, 2013;288(3):1953-66. Species: Human Sample Type: whole cells Application: Neut Isotretinoin increases skin-surface levels of neutrophil gelatinase-associated lipocalin in patients treated for severe acne. Authors: Lumsden KR, Nelson AM, Dispenza MC, Gilliland KL, Cong Z, Zaenglein AL, Thiboutot DM Br. J. Dermatol., 2011;165(2):302-10. Species: Human Sample Type: Whole Cells Application: Neut Soluble forms of Toll-like receptor 4 are present in human saliva and modulate tumour necrosis factor-alpha secretion by macrophage-like cells. Authors: Zunt SL, Burton LV, Goldblatt LI, Dobbins EE, Srinivasan M Clin. Exp. Immunol., 2009;156(2):285-93. Species: Human Sample Type: Saliva Application: WB
纯化方式	Antigen Affinity-purified
免疫原	Mouse myeloma cell line NS0-derived recombinant human TLR4 Glu24-Lys631 Accession # O00206
内毒素水平	<0.10 EU per 1 μg of the antibody by the LAL method.
生物活性	Human
标记	Unconjugated
溶解方法	Reconstitute at 0.2 mg/mL in sterile PBS.
背景	TLR4 is a 100 kDa type I transmembrane glycoprotein that belongs to the mammalian Toll-Like Receptor family of pathogen pattern recognition molecules. In the literature molecular weights correspondent to 110 kDa and 130 kDa were reported for TLR4 (1). MD-2, also known as ESOP-1, is a 25 kDa secreted protein that is required for TLR4-mediated responses to bacterial lipopolysaccharide (LPS) (1‑4). 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 (5). 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 (6‑8). MD-2 expression is required for cell surface localization of TLR4 and for optimal LPS-induced TLR4 signaling (8, 9). MD-2 also forms soluble disulfide-linked homo-oligomers which can interact with TLR4 (7). 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 (10‑12). The interaction of MD-2/LPS with TLR4 induces receptor oligomerization and the triggering of an inflammatory response (13). Increased levels of plasma MD-2 in septic shock patients sensitizes MD-2 non-expressing epithelial cells to LPS and promotes widespread tissue inflammation (14).
运输条件	Blue Ice
存放说明	4℃
参考文献	Ohnishi, T. et al. (2003) Clin. Diagn. Lab. Immouol. 10:405. Gangloff, M. and N.J. Gay (2004) Trends Biochem. Sci. 29:294. Palsson-McDermott, E.M. and L.A. O’Neill (2004) Immunology 113:153. Miyake, K. (2004) Semin. Immunol. 16:11. Medzhitov, R. et al. (1997) Nature 388:394. Shimazu, R. et al. (1999) J. Exp. Med. 189:1777. Visintin, A. et al. (2001) Proc. Natl. Acad. Sci. USA 98:12156. Akashi, S. et al. (2000) J. Immunol. 164:3471. Nagai, Y. et al. (2002) Nat. Immunol. 3:667. Re, F. and J.L. Strominger (2003) J. Immunol. 171:5272. Kennedy, M.N. et al. (2004) J. Biol. Chem. 279:34698. Gioannini, T.L. et al. (2004) Proc. Natl. Acad. Sci. USA 101:4186. Saitoh, S. et al. (2004) J. Endotoxin Res. 10:257. Pugin, J. et al. (2004) Blood 104:4071.

参考图片

Detection of Human TLR4 by Western Blot. Western blot shows lysates of human placenta tissue. PVDF membrane was probed with 1 µg/mL of Goat Anti-Human TLR4 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF1478) followed by HRP-conjugated Anti-Goat IgG Secondary Antibody (Catalog # HAF109). Specific bands were detected for TLR4 at approximately 110 kDa and 130 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 8.

TLR4 in Human PBMCs. TLR4 was detected in immersion fixed human peripheral blood mononuclear cells (PBMCs) using Goat Anti-Human TLR4 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF1478) at 10 µg/mL for 3 hours at room temperature. Cells were stained using the NorthernLights™ 557-conjugated Anti-Goat IgG Secondary Antibody (yellow; Catalog # NL001) and counterstained with DAPI (blue). View our protocol for Fluorescent ICC Staining of Non-adherent Cells.

TLR4 in Human Prostate. TLR4 was detected in immersion fixed paraffin-embedded sections of human prostate using Goat Anti-Human TLR4 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF1478) at 10 µg/mL overnight at 4 °C. Before incubation with the primary antibody tissue was subjected to heat-induced epitope retrieval using Antigen Retrieval Reagent-Basic (Catalog # CTS013). Tissue was stained using the Anti-Goat HRP-DAB Cell & Tissue Staining Kit (brown; Catalog # CTS008) and counterstained with hematoxylin (blue). View our protocol for Chromogenic IHC Staining of Paraffin-embedded Tissue Sections.

IL‑8 Secretion Induced by LPS and Neutralization by Human TLR4 Antibody. Lipopolysacharide (LPS) stimulates IL‑8 secretion in the HEK293 human embryonic kidney cell line co-transfected with human TLR4 and MD-2, in a dose-dependent manner (orange line), as measured by the Human CXCL8/IL‑8 Quantikine ELISA Kit (Catalog # D8000C). IL‑8 secretion elicited by LPS (75 ng/mL) is neutralized (green line) by increasing concentrations of Goat Anti-Human TLR4 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF1478). The ND₅₀ is typically 1.5-7.5 µg/mL.