Mouse Siglec-E Antibody

R&D Systems | Catalog # AF5806

R&D Systems

Key Product Details

Species Reactivity

Validated:

Mouse

Cited:

Human, Mouse, Hamster, Primate - Chlorocebus aethiops (African Green Monkey)

Applications

Validated:

Immunohistochemistry, Western Blot

Cited:

Immunohistochemistry-Paraffin, Western Blot, Neutralization, Immunocytochemistry, Immunoprecipitation, Functional Assay

Label

Unconjugated

Antibody Source

Polyclonal Goat IgG
View all Siglec-E Primary Antibodies »

Product Specifications

Immunogen

Mouse myeloma cell line NS0-derived recombinant mouse Siglec-E
Gln20-Phe355
Accession # Q6PJ50

Specificity

Detects mouse Siglec-E in direct ELISAs and Western blots. In direct ELISAs, less than 3% cross-reactivity with recombinant human (rh) Siglec-6, rhSiglec-7, and rhSiglec-9 is observed.

Clonality

Polyclonal

Host

Goat

Isotype

IgG

Scientific Data Images for Mouse Siglec-E Antibody

Detection of Mouse Siglec-E antibody by Western Blot.

Detection of Mouse Siglec‑E by Western Blot.

Western blot shows lysates of mouse spleen tissue. PVDF membrane was probed with 1 µg/mL of Goat Anti-Mouse Siglec-E Antigen Affinity-purified Polyclonal Antibody (Catalog # AF5806) followed by HRP-conjugated Anti-Goat IgG Secondary Antibody (Catalog # HAF109). A specific band was detected for Siglec-E at approximately 80 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.
Siglec-E antibody in Mouse Spleen by Immunohistochemistry (IHC-Fr).

Siglec‑E in Mouse Spleen.

Siglec-E was detected in perfusion fixed frozen sections of mouse spleen using Goat Anti-Mouse Siglec-E Antigen Affinity-purified Polyclonal Antibody (Catalog # AF5806) at 1.7 µg/mL overnight at 4 °C. Tissue was stained using the NorthernLights™ 557-conjugated Anti-Goat IgG Secondary Antibody (red; Catalog # NL001) and counterstained with DAPI (blue). Specific staining was localized to plasma membrane. View our protocol for Fluorescent IHC Staining of Frozen Tissue Sections.

Applications for Mouse Siglec-E Antibody

Application
Recommended Usage

Immunohistochemistry

5-15 µg/mL
Sample: Perfusion fixed frozen sections of mouse spleen

Western Blot

1 µg/mL
Sample: Mouse spleen tissue

Formulation, Preparation, and Storage

Purification

Antigen Affinity-purified

Reconstitution

Reconstitute at 0.2 mg/mL in sterile PBS. For liquid material, refer to CoA for concentration.

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.

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.

Calculators

The reconstitution calculator allows you to quickly calculate the volume of a reagent to reconstitute your vial. Simply enter the mass of reagent and the target concentration and the calculator will determine the rest.

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Background: Siglec-E

Siglecs are sialic acid specific I‑type lectins that are characterized by an extracellular domain (ECD) with an N‑terminal Ig‑like V‑set domain followed by varying numbers of Ig‑like C2‑set domains (1, 2). Mouse Siglec‑E, also known as Myeloid Inhibitory Siglec (MIS), is an 80 ‑ 85 kDa member of the CD33‑related subfamily of Siglecs. It consists of a 335 amino acid (aa) ECD with one Ig‑like V‑set domain and two Ig‑like C2‑set domains, a 21 aa transmembrane segment, and a 93 aa cytoplasmic domain that contains two immunoreceptor tyrosine‑based inhibitory motifs (ITIM) (3, 4). Rodent and primate Siglec gene families have significantly diverged, and Siglec‑9 is the most likely human ortholog of mouse Siglec‑E (1). Within the ECD, mouse Siglec‑E shares 56% and 80% aa sequence identity with human Siglec‑9 and rat Siglec‑E, respectively. Siglec‑E is expressed as a heavily N‑glycosylated disulfide‑linked homodimer and shows binding preference for disialic acids in the alpha 2‑8 linkage (3, 5). It is expressed on the surface of several hematopoietic cell types including neutrophils, NK cells, monocytes, peritoneal macrophages and B1 cells, and splenic myeloid dendritic cells and marginal zone B cells (5). Tyrosine phosphorylation of the cytoplasmic ITIMs mediates the association of Siglec‑E with the phosphatases SHP‑1 and SHP‑2 (3, 4). Siglec‑E is up‑regulated and additionally phosphorylated following cellular stimulation by a variety of TLR agonists (6). Siglec‑E signaling negatively regulates the LPS‑induced production of TNF‑ alpha and IL‑6 by macrophages (4, 6). Its up‑regulation in macrophages parallels the development of endotoxin tolerance (6). Siglec‑E recognition of sialylated determinants on virulent T. cruzi contributes to the suppression of dendritic cell IL‑12 p40 production (7).

References

  1. Varki, A. and T. Angata (2006) Glycobiology 16:1R.
  2. Crocker, P.R. et al. (2007) Nat. Rev. Immunol. 7:255.
  3. Yu, Z. et al. (2001) Biochem. J. 353:483.
  4. Ulyanova, T. et al. (2001) J. Biol. Chem. 276:14451.
  5. Zhang, J.Q. et al. (2004) Eur. J. Immunol. 34:1175.
  6. Boyd, C.R. et al. (2009) J. Immunol. 183:7703.
  7. Erdmann, H. et al. (2009) Cell. Microbiol. 11:1600.

Long Name

Sialic Acid Binding Ig-like Lectin E

Alternate Names

SiglecE

Entrez Gene IDs

83382 (Mouse)

Gene Symbol

SIGLECE

UniProt

Q6PJ50

Additional Siglec-E Products

Product Documents for Mouse Siglec-E Antibody

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Product Specific Notices for Mouse Siglec-E Antibody

For research use only

Citations for Mouse Siglec-E Antibody

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Protocols

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