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Two-color flow cytometric analysis of CD300LG (Nepmucin) expression on mouse peritoneal exudate cells. BALB/c mouse peritoneal exudate cells (PECs) were isolated 3 days post-stimulation by intraperitoneal injection of thioglycolate solution and were preincubated with Purified Rat Anti-Mouse CD16/CD32 antibody (Mouse BD Fc Block™) (Cat. No. 553141/553142). The cells were then stained with PE Rat Anti-Mouse F4/80 antibody (Cat. No. 565410) and either Alexa Fluor® 647 Rat IgG2a, κ Isotype Control (Cat. No. 557906; Left Plot) or Alexa Fluor® 647 Rat Anti-Mouse CD300LG (Nepmucin) antibody (Cat. No. 566997; Right Plot) at 0.5 µg/test. Bivariate pseudocolor density plots showing the correlated expression of CD300LG (Nepmucin) [or Ig Isotype control staining] versus F4/80 were derived from gated events with the forward and side light-scatter characteristics of total viable PECs. Flow cytometric analysis was performed using a BD FACSCelesta™ Flow Cytometer System and FlowJo™ software. Data shown on this Technical Data Sheet are not lot specific.
BD Pharmingen™ Alexa Fluor® 647 Rat Anti-Mouse CD300LG (Nepmucin)
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- Since applications vary, each investigator should titrate the reagent to obtain optimal results.
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- Alexa Fluor® 647 fluorochrome emission is collected at the same instrument settings as for allophycocyanin (APC).
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配套商品
The ZAQ2 monoclonal antibody specifically recognizes CD300LG, a microvascular endothelial adhesion molecule that belongs to the CD300 antigen like family. CD300LG is also known as Nepmucin (a not expressed in Peyer's patches mucin) and CMRF35-like molecule 9 (CLM-9). Four isoforms of this type I transmembrane glycoprotein have been described that are encoded by Cd300lg. The extracellular region of this sialomucin contains one IgV-like domain and one mucin-like domain followed by a transmembrane region and an intracellular tail. The ZAQ2 antibody recognizes the Ig domain of CD300LG (Nepmucin). This sialomucin is variably expressed on the luminal surfaces of endothelial cells lining small arterioles, venules and capillaries in many tissues. Although CD300LG (Nepmucin) protein is expressed by high endothelial venules (HEV) in lymph nodes, it is not expressed by Peyer's patches HEV. Nepmucin mRNA expression has also been observed for some myeloid lineage cell lines as well as CD11c+ dendritic cells, but not purified T and B lymphocytes. HEV-associated CD300LG (Nepmucin) mediates homotypic interactions between endothelial cells. This sialomucin also supports L-selectin-dependent lymphocyte rolling and heterophilic binding of lymphocytes by endothelial cells leading to transendothelial migration (TEM) of lymphocytes. The ZAQ2 antibody can reportedly inhibit lymphocyte TEM. CD300LG (Nepmucin) expression on microvascular endothelial cells is affected by local factors generated in tissues, including sites undergoing inflammation or tumor cell infiltration. This may in turn affect the levels of lymphocytic infiltration influenced by this sialomucin.
研发参考 (4)
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Jin S, Umemoto E, Tanaka T, et al. Nepmucin/CLM-9, an Ig domain-containing sialomucin in vascular endothelial cells, promotes lymphocyte transendothelial migration in vitro.. FEBS Lett. 2008; 582(20):3018-24. (Clone-specific: Functional assay, Immunofluorescence, Inhibition). 查看参考
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Umemoto E, Hayasaka H, Bai Z, et al. Novel regulators of lymphocyte trafficking across high endothelial venules.. Crit Rev Immunol. 2011; 31(2):147-69. (Biology). 查看参考
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Umemoto E, Takeda A, Jin S, et al. Dynamic changes in endothelial cell adhesion molecule nepmucin/CD300LG expression under physiological and pathological conditions.. PLoS ONE. 2013; 8(12):e83681. (Clone-specific: Immunofluorescence, Immunohistochemistry). 查看参考
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Umemoto E, Tanaka T, Kanda H, et al. Nepmucin, a novel HEV sialomucin, mediates L-selectin-dependent lymphocyte rolling and promotes lymphocyte adhesion under flow.. J Exp Med. 2006; 203(6):1603-14. (Immunogen: ELISA, Immunofluorescence, Immunohistochemistry, Immunoprecipitation, Western blot). 查看参考
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