Human Siglec-11 Antibody

Catalog #: AF3258 Datasheet / COA / SDS

Catalog #	Availability	Size / Price	Qty
AF3258
AF3258-SP

Detection of Siglec‑11 in Human PBMCs by Flow Cytometry.

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All Siglec-11 Antibodies

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Human Siglec-11 Antibody Summary

Species Reactivity

Human

Specificity

Detects human Siglec-11 in direct ELISAs and Western blots. In direct ELISA and Western blots, approximately 20% cross-reactivity with recombinant human (rh) Siglec-10 is observed and less than 5% cross-reactivity with rhSiglec-2, -3, -5, -6, -7, -9 and recombinant mouse Siglec-F is observed.

Source

Polyclonal Goat IgG

Purification

Antigen Affinity-purified

Immunogen

Chinese hamster ovary cell line CHO-derived recombinant human Siglec-11
Asn17-His543 (Glu84Ala, Lys145Gln)
Accession # Q96RL6

Formulation

Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. *Small pack size (SP) is supplied either lyophilized or as a 0.2 µm filtered solution in PBS.

Applications

Recommended Concentration

Sample

Western Blot

0.1 µg/mL

Recombinant Human Siglec‑11 Fc Chimera (Catalog # 3258-SL)

Flow Cytometry

2.5 µg/10⁶ cells

See below

Blockade of Receptor-ligand Interaction

In a functional ELISA, 1.5-4.5 µg/mL of this antibody will block 50% of the binding of 1 μg/mL of biotinylated (Neu5Ac alpha 2-8)₂-Polyacrylamide to immobilized Recombinant Human Siglec-11 Fc Chimera (Catalog # 3258-SL) coated at 2.5 µg/mL (100 µL/well). At 30 μg/mL, this antibody will block >90% of the binding.

CyTOF-ready

Ready to be labeled using established conjugation methods. No BSA or other carrier proteins that could interfere with conjugation.

Please Note: Optimal dilutions should be determined by each laboratory for each application. General Protocols are available in the Technical Information section on our website.

Scientific Data

Flow Cytometry

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Detection of Siglec‑11 in Human PBMCs by Flow Cytometry. Human peripheral blood monocytes (PBMC) were labeled with Goat Anti-Human Siglec-11 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF3258) followed by Phycoerythrin-conjugated Anti-Goat IgG Secondary Antibody (Catalog # F0107) and Human CD14 APC-conjugated Monoclonal Antibody (Catalog # FAB3832A). Quadrant markers were set based on control antibody staining (Catalog # AB-108-C).

Reconstitution Calculator

Preparation and Storage

Reconstitution

Reconstitute at 0.2 mg/mL in sterile PBS.

Shipping

Lyophilized product is shipped at ambient temperature. Liquid small pack size (-SP) is shipped with polar packs. Upon receipt, store immediately at the temperature recommended below.

Stability & Storage

Use a manual defrost freezer and avoid repeated freeze-thaw cycles.

12 months from date of receipt, -20 to -70 °C as supplied.
1 month, 2 to 8 °C under sterile conditions after reconstitution.
6 months, -20 to -70 °C under sterile conditions after reconstitution.

Background: Siglec-11

Siglecs (sialic acid binding Ig-like lectins) are I-type lectins that belong to the immunoglobulin superfamily. They are characterized by an N‑terminal Ig-like V-set domain which mediates sialic acid binding, followed by a varying numbers of Ig-like C2-set domains. Siglecs‑3 and 5‑13 constitute the CD33/Siglec-3 related group, which are defined by their sequence homology and differential expression in the ematopoietic system (1‑3). Mature human Siglec-11 consists of a 534 amino acid (aa) extracellular domain (ECD), a 23 aa transmembrane segment, and a 114 aa cytoplasmic domain. The ECD contains one Ig-like V-set domain, and three Ig-like C2-set domains. The cytoplasmic domain contains two immunoreceptor tyrosine-based inhibitory motifs (ITIMs) (4). A splice variant of Siglec-11 has a deletion of nearly 100 aa in the extracellular juxtamembrane region. Among siglecs, the ECD of Siglec-11 is most closely related to that of Siglec-10 (82% aa sequence identity). The cytoplasmic domains of these proteins are only 20% identical. Siglec-11 is closely related to the pseudogenes Siglec-14 and Siglec-16 (4, 5). Human Siglec-11 shares 90%‑96% aa sequence identity with Siglec-11 from great apes. Rodent orthologs of Siglec-11 have not been identified. In human, Siglec-11 is expressed in tissue macrophages, brain microglia, and inflammatory site monocytes (4). Strong microglial expression is specific to humans, as it is less prominent or absent in chimpanzees and orangutans (5). Siglec-11 forms 180 kDa disulfide-linked dimers. It shows a strong binding preference for sialic acid in alpha 2-8 linkage which is unusual for siglecs (4). A conserved arginine in the Ig-like V-set domain only partially contributes to Siglec-11 ligand recognition, in contrast to its being required in other siglecs (4). Tyrosine phosphorylation of the cytoplasmic region of Siglec-11 promotes association with the phosphatases SHP-1 and SHP-2 (4).

References