Invitrogen
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Description: The monoclonal antibody eBio440c recognizes Siglec-H, a protein exclusively found on pDC (plasmacytoid dendritic cells) or type I IFN-producing cells (IPC) in the naive mouse. mouse IPC are typically PDCA+, CD11c+, CD11b-, B220+, and Ly-6C+, and are quick to respond to viruses. Siglec-H is a transmembrane protein of the Ig superfamily that like CD33 have been shown to bind sialic acid but lacks the characteristic cytoplasmic ITIM domain (immunoreceptor tyrosine based inhibitory motif). To overcome the lack of a cytoplasmic domain, Siglec-H associates with DAP12 thereby allowing for signal transduction.
The eBio440c antibody has been shown to inhibit pDC function (inhbits IFNalpha secretion in response to CpG).
It has been observed that some mouse strains (such as SJL) have higher percentages of pDCs compared to C57BL/6.
Applications Reported: This eBio440c antibody has been reported for use in flow cytometric analysis.
Applications Tested: This eBio440c antibody has been tested by flow cytometric analysis of SJL spleen cells. This can be used at less than or equal to 0.125 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 (Product # 00-8222) (100 µL cell sample + 100 µL IC Fixation Buffer) or 1-step Fix/Lyse Solution (Product # 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.
Siglec-H, a protein exclusively found on pDC (plasmacytoid dendritic cells) or type I IFN-producing cells (IPC) in the naive mouse. Mouse IPC are typically PDCA+, CD11c+, CD11b-, B220+, and Ly-6C+, and are quick to respond to viruses. Siglec-H is a transmembrane protein of the Ig superfamily that like CD33 have been shown to bind sialic acid but lacks the characteristic cytoplasmic ITIM domain (immunoreceptor tyrosine based inhibitory motif). To overcome the lack of a cytoplasmic domain, Siglec-H associates with DAP12 thereby allowing for signal transduction. The eBio440c antibody has been shown to inhibit pDC function (inhbits IFNalpha secretion in response to CpG). It has been observed that some mouse strains (such as SJL) have higher percentages of pDCs compared to C57BL/6.
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.
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Protein Aliases: sialic acid binding Ig-like lectin H; Siglec; SIGLEC-like protein
Gene Aliases: 6430529G09Rik; Siglec-H
Entrez Gene ID: (Mouse) 233274
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