Preparation And Storage
Recommended Assay Procedures
The biotinylated HTA125 antibody can be used for the immunofluorescent staining along with Streptavidin-phycoerythrin (Cat. No. 554061) for flow cytometric analyses of human leukocytes and cell lines that express TLR4 (see Figure).
Product Notices
- This reagent has been pre-diluted for use at the recommended Volume per Test. We typically use 1 × 10^6 cells in a 100-µl experimental sample (a test).
- Since applications vary, each investigator should titrate the reagent to obtain optimal results.
- Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
- Source of all serum proteins is from USDA inspected abattoirs located in the United States.
- Caution: Sodium azide yields highly toxic hydrazoic acid under acidic conditions. Dilute azide compounds in running water before discarding to avoid accumulation of potentially explosive deposits in plumbing.
The HTA125 monoclonal antibody specifically binds to the human Toll-like receptor 4 (TLR4), also known as CD284. Toll-like receptors (TLR) are type I transmembrane proteins characterized by an extracellular domain containing leucine-rich repeats and a cytoplasmic domain similar to IL-1R family. In humans, ten TLR have been identified so far. The TLR family acts as a pattern-recognition receptor that is an essential component of pathogen recognition and innate immunity. TLR4 has been identified as the receptor for Gram-negative bacterial lipopolysaccharide (LPS). TLR4 mRNA expression has been found in spleen, placenta, ovary, intestine and lung. Using HTA125, TLR4 protein expression has been detected in monocytes, B cells and T cells in human peripheral blood. The human TLR4 gene has been mapped to chromosome 9q32. The immunogen used to generate the HTA125 hybridoma was human TLR4-transfected Ba/F3 cells. The HTA125 antibody reportedly blocks LPS-triggered, TLR4-mediated stimulation of cells.
Development References (10)
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Akashi S, Ogata H, Kirikae F, et al. Regulatory roles for CD14 and phosphatidylinositol in the signaling via toll-like receptor 4-MD-2. Biochem Biophys Res Commun. 2000; 268(1):172-177. (Clone-specific). View Reference
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Akira S. Toll-like receptors and innate immunity. Adv Immunol. 2001; 78:1-56. (Biology). View Reference
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Beutler B. Tlr4: central component of the sole mammalian LPS sensor. Curr Opin Immunol. 2000; 12(1):20-26. (Biology). View Reference
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Chuang T, Ulevitch RJ. Identification of hTLR10: a novel human Toll-like receptor preferentially expressed in immune cells. Biochim Biophys Acta. 2001; 1518(1-2):157-161. (Biology). View Reference
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Chuang TH, Ulevitch RJ. Cloning and characterization of a sub-family of human toll-like receptors: hTLR7, hTLR8 and hTLR9. Eur Cytokine Netw. 2000; 11(3):372-378. (Biology). View Reference
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Du X, Poltorak A, Wei Y, Beutler B. Three novel mammalian toll-like receptors: gene structure, expression, and evolution. Eur Cytokine Netw. 2000; 11(3):362-371. (Biology). View Reference
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Rock FL, Hardiman G, Timans JC, Kastelein RA, Bazan JF. A family of human receptors structurally related to Drosophila Toll. Proc Natl Acad Sci U S A. 1998; 95(2):588-593. (Biology). View Reference
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Shimazu R, Akashi S, Ogata H, et al. MD-2, a molecule that confers lipopolysaccharide responsiveness on Toll-like receptor 4. J Exp Med. 1999; 189(11):1777-1782. (Immunogen). View Reference
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Takeuchi O, Kawai T, Sanjo H, et al. TLR6: A novel member of an expanding toll-like receptor family. Gene. 1999; 231(1-2):59-65. (Biology). View Reference
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Zarember KA, Godowski PJ. Tissue expression of human Toll-like receptors and differential regulation of Toll-like receptor mRNAs in leukocytes in response to microbes, their products, and cytokines. J Immunol. 2002; 168(2):554-561. (Biology). View Reference
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Comparisons, where applicable, are made against older BD Technology, manual methods or are general performance claims. Comparisons are not made against non-BD technologies, unless otherwise noted.
For Research Use Only. Not for use in diagnostic or therapeutic procedures.