MAG/Siglec-4a Antibody (1099521) [Unconjugated]

Western Blot shows lysates of human cerebellum, mouse cerebellum and rat cerebellum. PVDF membrane was probed with 1 µg/ml of Mouse Anti-Human MAG/Siglec‑4a Monoclonal Antibody (Catalog # MAB11688) followed by HRP-conjugated Anti-Mouse IgG Secondary Antibody (Catalog # HAF018). A specific band was detected for MAG/Siglec‑4a at approximately 100 kDa (as indicated). This experiment was conducted under reducing conditions and using Western Blot Buffer Group 1.

MAG/Siglec‑4a was detected in immersion fixed paraffin-embedded sections of human cerebellum using Mouse Anti-Human MAG/Siglec‑4a Monoclonal Antibody (Catalog # MAB11688) at 5 µg/ml for 1 hour at room temperature followed by incubation with the Anti-Mouse IgG VisUCyte™ HRP Polymer Antibody (Catalog # VC001) or the HRP-conjugated Anti-Mouse IgG Secondary Antibody (Catalog # HAF007). Before incubation with the primary antibody, tissue was subjected to heat-induced epitope retrieval using VisUCyte Antigen Retrieval Reagent-Basic (Catalog # VCTS021). Tissue was stained using DAB (brown) and counterstained with hematoxylin (blue). Specific staining was localized to the membrane of processes. View our protocol for IHC Staining with VisUCyte HRP Polymer Detection Reagents.

MAG/Siglec‑4a was detected in immersion fixed paraffin-embedded sections of human brain cortex using Mouse Anti-Human MAG/Siglec‑4a Monoclonal Antibody (Catalog # MAB11688) at 5 µg/ml for 1 hour at room temperature followed by incubation with the Anti-Mouse IgG VisUCyte™ HRP Polymer Antibody (Catalog # VC001) or the HRP-conjugated Anti-Mouse IgG Secondary Antibody (Catalog # HAF007). Before incubation with the primary antibody, tissue was subjected to heat-induced epitope retrieval using VisUCyte Antigen Retrieval Reagent-Basic (Catalog # VCTS021). Tissue was stained using DAB (brown) and counterstained with hematoxylin (blue). Specific staining was localized to the membrane of processes. View our protocol for IHC Staining with VisUCyte HRP Polymer Detection Reagents.

• Reactivity: Hu
• Applications: WB, IHC
• Clone: 1099521
• Clonality: Monoclonal
• Host: Mouse
• Conjugate: Unconjugated

MAG/Siglec-4a Antibody (1099521) [Unconjugated] Summary

• Immunogen: Human embryonic kidney cell, HEK293-derived human MAG/Siglec-4a
  Gly20-Pro516
• Specificity: Detects recombinant human MAG protein in Direct ELISA.
• Source: N/A
• Isotype: IgG2b
• Clonality: Monoclonal
• Host: Mouse
• Purity Statement: Protein A or G purified from hybridoma culture supernatant

Applications/Dilutions

• Dilutions:
  • Immunohistochemistry 3-25 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.
• Reconstitution Instructions: Reconstitute lyophilized material at 0.2 mg/ml in sterile PBS. For liquid material, refer to CoA for concentration.

Notes

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

Alternate Names for MAG/Siglec-4a Antibody (1099521) [Unconjugated]

• GMAsialic acid binding Ig-like lectin 4A
• MAG
• myelin associated glycoprotein
• myelin-associated glycoprotein
• sialic acid-binding immunoglobulin-like lectin 4A
• Siglec4a
• Siglec-4a
• S-MAG

Background

Myelin-Associated Glycoprotein (MAG), also known as Siglec-4a, is a type I transmembrane glycoprotein belonging to the Siglec family, a subgroup of the Ig superfamily (1). It is composed of an extracellular segment containing five Ig-like domains, a single transmembrane segment, and a cytoplasmic domain. Mature MAG exists as two isoforms, termed S-MAG (short) and L-MAG (long), due to alternative splicing of the cytoplasmic domain (1, 2). S-MAG has a predicted molecular weight of 67 kDa while L-MAG has a predicted molecular weight of 71 kDa (1, 2). Additionally, proteolytic cleavage of the extracellular domain produces a soluble MAG (3). Within shared regions in the extracellular domain, human MAG shares 95% aa sequence identity with mouse and rat MAG. MAG functions as an adhesion molecule during neural development. It preferentially binds to alpha -2,3-linked sialic acid terminal structures found on cell surface molecules (1, 4, 5). MAG is selectively expressed by myelinating oligodendrocytes and Schwann cells and plays an important role in axon-myelin stability (1, 4). Specifically, L-MAG is involved in myelination in the central nervous system (CNS) while S-MAG is the predominate isoform expressed during myelination in the peripheral nervous system (1). MAG is also reported to regulate the axon cytoskeleton and support the distribution of axon molecules at the nodes of Ranvier (1, 4). In addition, it has been identified as a major inhibitor of neurite outgrowth (1, 4, 6). However, MAG has also been reported to protect neurons from excitotoxicity (1, 7). MAG is believed to utilize the gangliosides GD1a and GT1b, the Nogo receptors NgR1 and NgR2/NgRH1, Integrin beta 1/CD29, and PIR-B to mediate its effects (1, 4, 5, 8, 9). Soluble MAG, which is released from myelin in large quantities, has been identified in normal human tissues and in tissues from patients with neurological disorders (3). It is believed that this soluble MAG might contribute to the lack of CNS neuron regeneration after injury (3).

1. Lopez, P.H. (2014) Adv. Neurobiol. 9:245.
2. Salzer, J.L. et al. (1987) J. Cell Biol. 104:957.
3. Tang, S. et al. (1997) Mol. Cell. Neurosci. 9:333.
4. Schnaar, R.L. and P.H. Lopez (2009) J. Neurosci. Res. 87:3267.
5. Schnaar, R.L. (2010) FEBS Lett. 584:1741.
6. Akbik, F. et al. (2012) Exp. Neurol. 235:43.
7. Lopez, P.H. et al. (2011) J. Neurochem. 116:900.
8. Atwal, J.K. et al. (2008) Science 322:967.
9. Goh, E.L. et al. (2008) Mol. Brain 1:10.

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.