Each lot of this antibody is quality control tested by immunofluorescent staining with flow cytometric analysis. For immunofluorescent staining, the suggested use of this reagent is ≤ 0.5 µg per 106 cells in 100 µl volume or per 100 µl whole blood. It is recommended that the reagent be titrated for optimal performance for each application.
Application Notes:
Additional reported applications (for the relevant formats) include: immunohistochemical staining4 of acetone-fixed frozen sections, immunofluorescence microscopy6, and in vitro blocking of LPS-induced cytokine production2,3,7.9. For most successful immunofluorescent staining results, it may be important to maximize signal over background by using a relatively bright fluorochrome-antibody conjugate (Cat. No. 312806) or by using a high sensitivity, three-layer staining technique (e.g., including a biotinylated antibody (Cat. No. 312804) or biotinylated anti-mouse IgG second step (Cat. No. 405303), followed by SAv-PE (Cat. No. 405204)). The LEAF™ purified antibody (Endotoxin <0.1 EU/μg, Azide-Free, 0.2 μm sterile-filtered) is recommended for functional assays (Cat. No. 312807).
Application References:
1. Skimazu, R., et al., 1999. J. Exp. Med. 189 :1777. 2. Wang, R., et al., 2003. Hybrid Hybridomics 22 :357. 3. Wang, J.E., et al., Infect. Immun. 69 :2402. 4. Ishihara, S., et al., 2004 J. Immunol. 173 :1406. 5. Kawahara, T., et al., 2001 Infec. Immun 69:4382. 6. Jiang, Q. et al., 2000. J. Immunol. 165:3541. 7. Sugawara, S., et al., 2001. Infect. Immun. 69:4951. 8. Chavakis, E., et al., 2007. Circ. Res. 100:204.PubMed 9. Bhattacharyya, S., et al., 2007. Am. J. Physiol. Gastrointest Liver Physiol. doi:10.1152/ajpgi.00149.PubMed
Human peripheral blood monocytes stained with HTA125 PE
Description:
Toll-like receptors are type I transmembrane signaling receptors. They are primordial pathogen-recognition proteins that function as sentinels for the innate immune system. TLR4, also known as CD284, is a 110 kD protein which is expressed on monocytes/macrophages, endothelial cells, and at low levels on B cells and granulocytes. In association with a secretory molecule, MD2, TLR4 has been recognized as critical for host recognition of bacterial LPS. HTA125 antibody is useful for flow cytometric analysis and is able to block LPS-induced cytokine production.
Other Names:
Toll-like Receptor 4, TLR4, TLR-4, CD284
Structure:
Type I transmembrane protein, Toll-like receptor family/IL-1R superfamily, 110 kD
Function:
Activates the innate immune system against Gram-negative bacterial pathogens by recognition of LPS
Ligand Receptor:
Bacterial LPS
Antigen References:
1. Skimazu, R., et al., 1999. J. Exp. Med. 189 :1777.
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