|Tested species reactivity||Human|
|Published species reactivity||Not Applicable|
|Host / Isotype||Mouse / IgG2a, kappa|
|Storage buffer||PBS, pH 7.2, with 0.1% gelatin, 0.2% BSA|
|Contains||0.09% sodium azide|
|Storage Conditions||4° C, store in dark, DO NOT FREEZE!|
|Tested Applications||Dilution *|
|Flow Cytometry (Flow)||5 µL (1 µg)/test|
* Suggested working dilutions are given as a guide only. It is recommended that the user titrate the product for use in their own experiment using appropriate negative and positive controls.
Description: The HTA125 monoclonal antibody reacts with human Toll-like receptor 4 (TLR4). So far, at least ten members of the Toll family have been identified in humans. This family of type I transmembrane proteins is characterized by an extracellular domain with leucine-rich repeats and a cytoplasmic domain with homology to the type I IL-1 receptor. Two of these receptors, TLR2 and TLR4, are pattern recognition receptors and signaling molecules in response to bacterial lipoproteins and have been implicated in innate immunity and inflammation. TLR4 physically associates with another molecule called MD-2, and together with CD14, this complex is responsible for LPS recognition and signaling. TLR4 is expressed by peripheral blood monocytes. HTA125 has been reported to immunoprecipitate human TLR4 (~100 kDa) from transfected cells. Most TLR cell surface expression, especially TLR1 and TLR4, occurs at low levels on monocytes and at even lower levels on other cell types including granulocytes and immature dendritic cells (iDC). Furthermore, a relatively high degree of variability in TLR surface expression has been reported among normal donors.
Applications Reported: This HTA125 antibody has been reported for use in flow cytometric analysis.
Applications Tested: This HTA125 antibody has been pre-titrated and tested by flow cytometric analysis of normal human peripheral blood cells. This can be used at 5 µL (1 µg) per test. A test is defined as the amount (µg) of antibody that will stain a cell sample in a final volume of 100 µL. Cell number should be determined empirically but can range from 10^5 to 10^8 cells/test.
It is highly recommended that for optimal staining of TLR4, whole blood be stained using the LWB protocol rather than Ficoll-gradient prepared normal human peripheral blood cells. The use of a density gradient appears to reduce the staining intensity significantly.
Light sensitivity: This tandem dye is sensitive photo-induced oxidation. Please protect this vial and stained samples from light.
Fixation: Samples can be stored in IC Fixation Buffer (cat. 00-8222) (100 µL cell sample + 100 µL IC Fixation Buffer) or 1-step Fix/Lyse Solution (cat. 00-5333) for up to 3 days in the dark at 4°C with minimal impact on brightness and FRET efficiency/compensation. Some generalizations regarding fluorophore performance after fixation can be made, but clone specific performance should be determined empirically.
Excitation: 488-561 nm; Emission: 775 nm; Laser: Blue Laser, Green Laser, Yellow-Green Laser
TLR4 is a member of the Toll like receptor (TLR) family which plays a fundamental role in pathogen recognition and activation of innate immunity. TLRs are highly conserved from Drosophila to humans and share structural and functional similarities. This receptor is most abundantly expressed in placenta, and in myelomonocytic subpopulation of the leukocytes. It has been implicated in signal transduction events induced by lipopolysaccharide (LPS) found in most gram-negative bacteria.
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.
|Not Applicable||Not Cited||Human heat shock protein 60 stimulates vascular smooth muscle cell proliferation through Toll-like receptors 2 and 4.||de Graaf R,Kloppenburg G,Kitslaar PJ,Bruggeman CA,Stassen F||Microbes and infection (8:1859)||2006|
|Not Applicable||Not Cited||Expression and involvement of Toll-like receptors (TLR)2, TLR4, and CD14 in monocyte TNF-alpha production induced by lipopolysaccharides from Neisseria meningitidis.||Mirlashari MR,Lyberg T||Medical science monitor : international medical journal of experimental and clinical research (9:BR316)||2003|
|Not Applicable||Not Cited||Toll-like receptors confer responsiveness to lipopolysaccharide from Porphyromonas gingivalis in human gingival fibroblasts.||Tabeta K,Yamazaki K,Akashi S,Miyake K,Kumada H,Umemoto T,Yoshie H||Infection and immunity (68:3731)||2000|
|Not Applicable||Not Cited||Regulatory roles for CD14 and phosphatidylinositol in the signaling via toll-like receptor 4-MD-2.||Akashi S,Ogata H,Kirikae F,Kirikae T,Kawasaki K,Nishijima M,Shimazu R,Nagai Y,Fukudome K,Kimoto M,Miyake K||Biochemical and biophysical research communications (268:172)||2000|
|Not Applicable||Not Cited||MD-2, a molecule that confers lipopolysaccharide responsiveness on Toll-like receptor 4.||Shimazu R,Akashi S,Ogata H,Nagai Y,Fukudome K,Miyake K,Kimoto M||The Journal of experimental medicine (189:1777)||1999|