Description: The Sa2-8 monoclonal antibody reacts with mouse CD14, a 53-55 kDa GPI-linked glycoprotein. CD14 is a receptor for the complexes of LPS and LBP (LPS-Binding Protein) and is shown to associate with Toll-Like Receptor 4 (TLR4) and participate in the signaling and cellular response to bacterial LPS. In mouse, CD14 is expressed on the surface of macrophages and under certain conditions is also found in the serum in a secreted form. Sa2-8 has weak antagonistic activity (in NF-kappaB activation or TNF alpha production with LPS stimulation). Applications Reported: This Sa2-8 antibody has been reported for use in flow cytometric analysis. Applications Tested: This Sa2-8 antibody has been tested by flow cytometric analysis of mouse thicoglycolate-elicited peritoneal cells. This can be used at less than or equal to 0.5 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 recommended that the antibody be carefully titrated for optimal performance in the assay of interest. PerCP-eFluor® 710 emits at 710 nm and is excited with the blue laser (488 nm); it can be used in place of PerCP-Cyanine5.5. We recommend using a 710/50 bandpass filter, however, the 695/40 bandpass filter is an acceptable alternative.Please make sure that your instrument is capable of detecting this fluorochrome. 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 nm; Emission: 710 nm; Laser: Blue Laser. Filtration: 0.2 µm post-manufacturing filtered. CD14 is a 55 kDa GPI-anchored glycoprotein that is constitutively expressed on the surface of mature monocytes, macrophages, and neutrophils. CD14 also serves as a multifunctional lipopolysaccharide receptor, and is released to the serum both as a secreted and enzymatically cleaved GPI-anchored form. CD14 binds lipopolysaccharide molecule in a reaction catalyzed by lipopolysaccharide-binding protein (LBP), an acute phase serum protein. The soluble sCD14 can discriminate slight structural differences between lipopolysaccharides and is important for neutralization of serum allochthonous lipopolysaccharides by reconstituted lipoprotein particles. Further, CD14 has been shown to bind apoptotic cells, and can affect allergic, inflammatory and infectious processes. Alternative splicing results in multiple transcript variants encoding the same CD14 isoform. Diseases associated with CD14 dysfunction include mycobacterium chelonae infection and Croup.
|Store at 2-8°C. Do not freeze. Light-sensitive material.|
|PBS with 0.1% gelatin and 0.09% sodium azide; pH 7.2|
For Research Use Only.
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