Sialic acid binding Ig like 10

Siglec (sialic acid binding Ig like) are members of Ig superfamily expressed on cell surface, structurally they have a characteristic-terminal V-set Ig-like domain containing sialic acid binding site followed by varying number of C2-set domain (1).   Eleven functional members of the Siglec family have been identified so far. Most  Siglecs have one or more immune receptor tyrosine based inhibitory motifs (ITIMs) in their cytoplasmic tails implicated in negative regulatory signaling function.  In general the immunoglobulin domain family; members are components of immunoglobulins, neuroglia, cell surface glycoproteins, such as, T-cell receptors, CD2, CD4, CD8, and membrane glycoproteins, such as, butyrophilin and chondroitin sulfate proteoglycan core.  The pattern recognition receptors which recognizes pathogens or component of injured cells trigger activation of innate immune response. In association with CD24 the Siglec10 in humans and Siglec-G in mice protects the host against lethal response to pathological cell death and discriminates injured cell component from pathogen associated molecular patterns (2).    

Siglec10 is an immune system restricted membrane bound protein that is highly expressed in peripheral blood leukocytes.  Several variant of Siglec10 are cloned (Siglec10Sv2, SV4 and Sv3), Siglec10Sv3 that lack part of the exon 3 but keeps the reading frame and interaction site with Sias and the motif for the intracellular signaling (3). Siglec10 is localized within the cluster of genes on chromosome 19q13.3-4 that codes other Siglec family members. The first Siglecs to be identified are /sialoadhesion/Siglec1, CD22/Siglec2, CD33/Siglec3 and  myelin-associated glycoprotein (MAG)/Siglec4.  Later new Siglecs (Siglec5, 6, 7, 8, 9 and 10) were identified each have distinct sialic acid binding properties. With the exception of siglec6 which is expressed in placenta, all other Siglecs are expressed in discrete subsets of hemopoetic cells.  The cytoplasmic tail of Siglec10 contains 4 tyrosines, two of them are embedded in ITIM-signaling motifs (Y597 and Y667), both are phosphorylated by tyrosine kinases and are involved in intracellular signaling (3). Siglec10 is expressed on esonophils, leukocytes and B-cells, Siglec10 us found on minor subset of NK-like cells that express CD16 low affinity Fc receptor but lack the CD56 antigen.  Siglec10 appears to be inhibitory receptor for innate immune system.

 The Siglec10-selective antibody was generated against a peptide from human Siglec10 protein corresponding to amino acids 675-697.  The affinity purified mono epitope-specific rabbit polyclonal antibodies were purified on immobilized antigen based affinity chromatography.  The antibodies labels a broad band at 72-74kDa inSiglec10 western blot control samples.  The Siglect10 antibodies were conjugated to FITC and Biotin and  these conjugates are stabilized for long-term storage with  out significant loss of activity.  Limited quantities of antigenic blocking peptide and western blot positive controls samples are also available (please inquire for availability).  FabGennix has a wide range of antibodies to innate immune and other targets, for a complete listing please visit www. FabGennix.com

Immunogen:            Synthetic peptide corresponding to amino acids 675-697 of the human Siglec10 protein corresponding to amino acid sequence (rpr pe a rmp kgt qad yae vkf q) ,  the peptide was amidated and conjugated to a carrier protein for preparation of antigen to generate antibodies in rabbit.  

Concentration:        Siglec10-101AP, FITC-Siglec10 and Biotin-Siglec10 IgG concentrations 0.56-                               0.60 mg/ml. 

Applications:      Antibody Siglec10-101AP is ideal for WB, use of this antibody in other applications has not yet worked out.  The dilution of Siglec10-101AP antibody is for reference only, investigators are expected to determine the optimal conditions for specific assay.  WB;  1:500; IMM & i.p pull-down assays:> n.d.                                     

Reactivity:          This antibody detects a broad band of 74-76kDa and a smaller band of 42-44kDa in PC-Siglec10 samples.  The antibody also reacts to light band of 53kDa in PC-Siglec10 samples, the identity of this band is not known.

Protocols:                Standard protocol for various applications (WB, IMM and IHC) of this antibody is provided with the product specification sheet, however, FabGennix Int. Inc.

Form/Storage:         The antiserum is supplied in antibody stabilization buffer with 0.02% sodium azide.  For long-term storage of antibodies, store at -20^oC.  FabGennix Int. Inc. does not recommend storage of very dilute antibody solutions unless they are prepared in specially formulated multi use antibody dilution buffer (Cat # DilUbuffer).  Working solutions of antibodies in DilUbuffer should be filtered through 0.45ƒÝ filter after every use for long-term storage.

 

 

For research use only, not for diagnostic or therapeutic use.