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 mouse splenocytes. This may be used at less than or equal to 0.25 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.

Super Bright 600 is a tandem dye that can be excited with the violet laser line (405 nm) and emits at 600 nm. We recommend using a 610/20 bandpass filter. Please make sure that your instrument is capable of detecting this fluorochrome.

When using two or more Super Bright dye-conjugated antibodies in a staining panel, it is recommended to use Super Bright Complete Staining Buffer (Product # SB-4401) to minimize any non-specific polymer interactions. Please refer to the datasheet for Super Bright Staining Buffer for more information.

Light sensitivity: This tandem dye is sensitive to photo-induced oxidation. Please protect this vial and stained samples from light.

Fixation: Samples can be stored in IC Fixation Buffer (Product # 00-8222-49) (100 µL of cell sample + 100 µL of IC Fixation Buffer) or 1-step Fix/Lyse Solution (Product # 00-5333-57) 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: 405 nm; Emission: 600 nm; Laser: Violet Laser

Super Bright Polymer Dyes are sold under license from Becton, Dickinson and Company.