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Mouse Siglec-E Antibody

Catalog #: AF5806
9 Citations Datasheet / COA / SDS
Catalog # Availability Size / Price Qty
AF5806
AF5806-SP
Detection of Mouse Siglec‑E by Western Blot.
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Product Details
Citations (9)
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Summary Data Examples Preparation and Storage Reconstitution Calculator Background Product Datasheets Related Research Areas

Mouse Siglec-E Antibody Summary

Species Reactivity
Mouse
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.
Source
Polyclonal Goat IgG
Purification
Antigen Affinity-purified
Immunogen
Mouse myeloma cell line NS0-derived recombinant mouse Siglec-E
Gln20-Phe355
Accession # Q6PJ50
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.
Label
Unconjugated

Applications

Recommended Concentration
Sample
Western Blot
1 µg/mL
See below
Immunohistochemistry
5-15 µg/mL
See below

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

Western Blot Detection of Mouse Siglec-E antibody by Western Blot. View Larger

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.

Immunohistochemistry Siglec-E antibody in Mouse Spleen by Immunohistochemistry (IHC-Fr). View Larger

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.

Reconstitution Calculator

Reconstitution Calculator

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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Preparation and Storage

Reconstitution
Reconstitute at 0.2 mg/mL in sterile PBS.
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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-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
Entrez Gene IDs
83382 (Mouse)
Alternate Names
CD170;CD33L2;OBBP2;OB-BP2;Sialic acid-binding Ig-like lectin 5;SIGLEC5;SIGLEC-5; SiglecE; Siglec-E

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Citations for Mouse Siglec-E Antibody

R&D Systems personnel manually curate a database that contains references using R&D Systems products. The data collected includes not only links to publications in PubMed, but also provides information about sample types, species, and experimental conditions.

9 Citations: Showing 1 - 9
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  1. CD24-Siglec axis is an innate immune checkpoint against metaflammation and metabolic disorder.
    Authors: Xu W, Mingyue L, Jifeng Z et al.
    Cell Metab.
  2. Paired Siglec receptors generate opposite inflammatory responses to a human‐specific pathogen
    Authors: Flavio Schwarz, Corinna S Landig, Shoib Siddiqui, Ismael Secundino, Joshua Olson, Nissi Varki et al.
    The EMBO Journal
  3. Innate extracellular Hsp70 inflammatory properties are mediated by the interaction of Siglec-E and LOX-1 receptors
    Authors: Borges, TJ;Lima, K;Murshid, A;Lape, IT;Zhao, Y;Rigo, MM;Lang, BJ;Siddiqui, SS;Hui, E;Riella, LV;Bonorino, C;Calderwood, SK;
    bioRxiv : the preprint server for biology
    Species: Mouse, Primate - Chlorocebus aethiops (African Green Monkey)
    Sample Types: Cell Lysates, Whole Cells, Transfected Whole Cells
    Applications: Immunoprecipitation, Western Blot, Neutralization, Immunocytochemistry
  4. CD24-Siglec axis is an innate immune checkpoint against metaflammation and metabolic disorder.
    Authors: Xu W, Mingyue L, Jifeng Z et al.
    Cell Metab.
  5. Siglec-7 restores ?-cell function and survival and reduces inflammation in pancreatic islets from patients with diabetes
    Authors: G Dharmadhik, K Stolz, M Hauke, NG Morgan, A Varki, E de Koning, S Kelm, K Maedler
    Sci Rep, 2017-04-05;7(0):45319.
    Species: Human
    Sample Types: Whole Cells
    Applications: Functional Assay
  6. Studies on the detection, expression, glycosylation, dimerization and ligand binding properties of mouse Siglec-E
    Authors: Shoib Siddiqui
    J. Biol. Chem, 2016-12-05;0(0):.
    Species: Hamster, Mouse
    Sample Types: Whole Cells
  7. Leishmania donovani Utilize Sialic Acids for Binding and Phagocytosis in the Macrophages through Selective Utilization of Siglecs and Impair the Innate Immune Arm
    PLoS Negl Trop Dis, 2016-08-05;10(8):e0004904.
    Species: Mouse
    Sample Types: Cell Lysates
    Applications: Western Blot
  8. Siglec receptors impact mammalian lifespan by modulating oxidative stress.
    Authors: Schwarz F, Pearce O, Wang X, Samraj A, Laubli H, Garcia J, Lin H, Fu X, Garcia-Bingman A, Secrest P, Romanoski C, Heyser C, Glass C, Hazen S, Varki N, Varki A, Gagneux P
    Elife, 2015-04-07;4(0):.
    Species: Mouse
    Sample Types: Whole Tissue
    Applications: IHC-P
  9. Group B Streptococcus engages an inhibitory Siglec through sialic acid mimicry to blunt innate immune and inflammatory responses in vivo.
    Authors: Chang Y, Olson J, Beasley F, Tung C, Zhang J, Crocker P, Varki A, Nizet V
    PLoS Pathog, 2014-01-02;10(1):e1003846.
    Species: Mouse
    Sample Types: Cell Lysates
    Applications: Western Blot

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