Product Information

Contents: Alexa Fluor® 647 anti-mouse siglec H - marker for IPC/pDC
Catalog Number: 51-0333
Sizes: 25 ug, 100 ug
Formulation: Phosphate buffered saline, pH 7.2, less than or equal to 0.09% NaN₃, BSA and/or gelatin for protein stabilization
Storage Conditions: Store at 4°C.
DO NOT FREEZE.
LIGHT-SENSITIVE MATERIAL.
Clone: eBio440c
Isotype: Rat IgG2b

Staining of 129 splenocytes with FITC anti-mo/hu B220 (RA3-6B2) (cat. 11-0452) and 0.125 μg of Alexa Fluor® 647 Rat IgG2b Isotype Control (cat. 51-4031) (left) or 0.125 μg of Alexa Fluor® 647 anti-mouse siglec H/ pDC plasmacytoid DC (eBio440c) (right). Cells in the larger FSC gate were used for analysis.

Available Formats of This Product
Cat. No.	Format	Excite (nm)	Emit (nm)	Reported Applications
14-0333	Affinity Purified anti-mouse siglec H - marker for IPC/pDC plasmacytoid DC	N/A	N/A	FC IHC
11-0333	Fluorescein isothiocyanate (FITC) anti-mouse siglec H - marker for IPC/pDC plasmacytoid DC	488	518	FC
51-0333	Alexa Fluor® 647 anti-mouse siglec H - marker for IPC/pDC plasmacytoid DC	633	668	FC
Flow Cytometry Product Notes: Test Sizes: To accommodate multicolor flow cytometry, eBioscience is in the process of reducing test size volumes from 20 µl to 5 µl. Please check your antibody vial for the recommended test size. Fluorochrome Replacements: eBioscience is in the process of replacing all Pacific Blue® and APC-Alexa Fluor® 750 conjugated products with eFluor™ 450 and APC-eFluor™ 780 conjugated products, respectively.

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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 naïve 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

For research use only, not for diagnostic or therapeutic use. 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 splenocytes. This can be used at less than or equal to 0.25 μg per million cells in a 100 μl total staining volume. It is recommended that the antibody be carefully titrated for optimal performance in the assay of interest.

References

Amanda Blasius, William Vermi, Anne Krug, Fabio Facchetti, Marina Cella, and Marco Colonna. A cell-surface molecule selectively expressed on murine natural interferonα producing cells that blocks secretion of interferonα. Blood. 2004;103:4201-4206 (440c, FC, IH/F, FA PubMed)

Blasius AL, Giurisato E, Cella M, Schreiber RD, Shaw AS, Colonna M. Bone marrow stromal cell antigen 2 is a specific marker of type I IFN-producing cells in the naive mouse, but a promiscuous cell surface antigen following IFN stimulation. J Immunol. 2006 Sep 1;177(5):3260-5.(440c, FC, PubMed)

Blasius AL, Cella M, Maldonado J, Takai T, Colonna M. Siglec-H is an IPC-specific receptor that modulates type I IFN secretion through DAP12. Blood. 2006 Mar 15;107(6):2474-6. Epub 2005 Nov 17.(440c, FC, PubMed)

Kreisel FH, Blasius A, Kreisel D, Colonna M, Cella M. Interferon-producing cells develop from murine CD31(high)/Ly6C(-) marrow progenitors. Cell Immunol. 2006 Aug;242(2):91-8.

Related Products

Cat. 51-4031 Alexa Fluor® 647 Rat IgG2b Isotype Control

Pat. No. pending

Alexa Fluor® and Pacific Blue® are registered trademarks of and licensed under patents assigned to Molecular Probes, Inc. for research use only. This product is subject to an agreement between Molecular Probes, Inc. and eBioscience, and the manufacture, use, sale or import of this product may be subject to one or more U.S. patents, pending applications and corresponding foreign equivalents, owned by Molecular Probes, Inc. (a wholly owned subsidiary of Invitrogen Corp). The purchase of this product conveys to the buyer the non-transferable right to use the purchased amount of the product for life science research or as an ASR. The buyer cannot use this product for manufacturing or for any other screening (specifically including use in combination with microarrays or High Content Screening) or testing purpose, other than as an ASR. For information on purchasing a license to this product for purposes other than life science research or use as an ASR, contact Molecular Probes, Inc.