Siglec-E Antibody [Unconjugated]

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 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.

• Reactivity: Mu
• Applications: WB, IHC
• Clonality: Polyclonal
• Host: Goat
• Conjugate: Unconjugated
• Concentration: LYOPH

Siglec-E Antibody [Unconjugated] Summary

• 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.
• Source: N/A
• Isotype: IgG
• Clonality: Polyclonal
• Host: Goat
• Gene: Siglece
• Purity Statement: Antigen Affinity-purified

Applications/Dilutions

• Dilutions:
  • Immunohistochemistry 5-15 ug/mL
  • Western Blot 1 ug/mL

Packaging, Storage & Formulations

• 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.
• Buffer: 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.
• Preservative: No Preservative
• Concentration: LYOPH
• Reconstitution Instructions: Reconstitute at 0.2 mg/mL in sterile PBS.

Notes

This product is produced by and ships from R&D Systems, Inc., a Bio-Techne brand.

Alternate Names for Siglec-E Antibody [Unconjugated]

• CD170;CD33L2;OBBP2;OB-BP2;Sialic acid-binding Ig-like lectin 5;SIGLEC5;SIGLEC-5
• SiglecE
• Siglec-E

Background

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).

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.

Limitations

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Primary Antibodies are guaranteed for 1 year from date of receipt.

Publications for Siglec-E Antibody (AF5806)(10)

Publications using AF5806

• He, J;Zhang, F;Wu, B;Yu, W; ST8SIA6 Sialylates CD24 to Enhance Its Membrane Localization in BRCA Cells 2024-12-26 [PMID: 39791710] (Immunocytochemistry, Mouse)
• Borges, TJ;Lima, K;Murshid, A;Lape, IT;Zhao, Y;Rigo, MM;Lang, BJ;Siddiqui, SS;Hui, E;Riella, LV;Bonorino, C;Calderwood, SK; Innate extracellular Hsp70 inflammatory properties are mediated by the interaction of Siglec-E and LOX-1 receptors bioRxiv : the preprint server for biology 2023-12-04 [PMID: 38106019] (Immunoprecipitation, Western Blot, Neutralization, Immunocytochemistry, Mouse, Primate - Chlorocebus aethiops (African Green Monkey))
• Xu W, Mingyue L, Jifeng Z et al. CD24-Siglec axis is an innate immune checkpoint against metaflammation and metabolic disorder. Cell Metab. 8/2/2022 [PMID: 35921817]
• Xu W, Mingyue L, Jifeng Z et al. CD24-Siglec axis is an innate immune checkpoint against metaflammation and metabolic disorder. Cell Metab. 2022-08-02 [PMID: 35921817]
• G Dharmadhik, K Stolz, M Hauke, NG Morgan, A Varki, E de Koning, S Kelm, K Maedler Siglec-7 restores ?-cell function and survival and reduces inflammation in pancreatic islets from patients with diabetes Sci Rep, 2017-04-05;7(0):45319. 2017-04-05 [PMID: 28378743] (Functional Assay, Human)
• Flavio Schwarz, Corinna S Landig, Shoib Siddiqui, Ismael Secundino, Joshua Olson, Nissi Varki et al. Paired Siglec receptors generate opposite inflammatory responses to a human‐specific pathogen The EMBO Journal 3/15/2017 [PMID: 28100677]
• Shoib Siddiqui Studies on the detection, expression, glycosylation, dimerization and ligand binding properties of mouse Siglec-E J. Biol. Chem, 2016-12-05;0(0):. 2016-12-05 [PMID: 27920204] (Hamster, Mouse)
• 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. 2016-08-05 [PMID: 27494323] (Western Blot, Mouse)
• 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 Siglec receptors impact mammalian lifespan by modulating oxidative stress. Elife, 2015-04-07;4(0):. 2015-04-07 [PMID: 25846707] (IHC-P, Mouse)
• Chang Y, Olson J, Beasley F, Tung C, Zhang J, Crocker P, Varki A, Nizet V Group B Streptococcus engages an inhibitory Siglec through sialic acid mimicry to blunt innate immune and inflammatory responses in vivo. PLoS Pathog, 2014-01-02;10(1):e1003846. 2014-01-02 [PMID: 24391502] (Western Blot, Mouse)

Bioinformatics

• Gene Symbol: Siglece
• Uniprot:
  • Q6PJ50()