Human MAG/Siglec-4a Antibody Summary
Gly20-Pro516
Applications
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
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Detection of Human, Mouse and Rat MAG/Siglec‑4a by Western Blot. 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.
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Detection of MAG/Siglec‑4a in Human Cerebellum. 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.
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Detection of MAG/Siglec‑4a in Human Brain Cortex. 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.
Reconstitution Calculator
Preparation and Storage
- 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: MAG/Siglec-4a
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).
- Lopez, P.H. (2014) Adv. Neurobiol. 9:245.
- Salzer, J.L. et al. (1987) J. Cell Biol. 104:957.
- Tang, S. et al. (1997) Mol. Cell. Neurosci. 9:333.
- Schnaar, R.L. and P.H. Lopez (2009) J. Neurosci. Res. 87:3267.
- Schnaar, R.L. (2010) FEBS Lett. 584:1741.
- Akbik, F. et al. (2012) Exp. Neurol. 235:43.
- Lopez, P.H. et al. (2011) J. Neurochem. 116:900.
- Atwal, J.K. et al. (2008) Science 322:967.
- Goh, E.L. et al. (2008) Mol. Brain 1:10.
Product Datasheets
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