Anti-Poly-Neu5Ac Antibody, clone 12E3

Cell surface of vertebrate cells contains a dense layer of glycans that often terminate with sialic acids, mainly Neu5Ac or Neu5Gc that differ only by a single oxygen atom. Glycoproteins and proteoglycans containing these moieties provide a scaffold for cell-to-cell and cell to environment communication. These sialic acids are mainly present as monosialyl residues at the non-reducing terminus of glycoproteins and play a role in ligand-receptor interactions during fertilization, development, differentiation, and adult brain functions. However, occasionally, sialic acids are linked to each other to form a polymerized structure (poly-sia; poly-Neu5Ac) that can contain 8 to 200 sialyl residues. These glycans are not only modifications of core molecules that are helpful as markers on particular cells, but also have independent functions via their interactions with glycan-recognizing molecules, such as lectins. Neu5Ac is reported to be a prevalent on a variety of sialoglycoconjugates SGCs) in vertebrate brain tissue. Its homopolymeric form (Poly-Neu5Ac) is shown to be most prevalent on the neural cell adhesion molecule (NCAM) and in the capsules of certain neuroinvasive bacteria. NCAM with polymeric Neu5Ac is highly expressed during the embryonic and early postnatal development and plays a critical role in neurite outgrowth and neural plasticity. Poly-Neu5Ac also acts as a neurotropic factor in neuroinvasive E. coli K1 and Neisseria meningitidis serogroup B. The exact mechanisms for breakdown of polymeric forms is not well understood. However, it can undergo an intramolecular self-cleavage into short oligomers under prolonged mild acidic conditions found in lysosomes. Clone 12E3 is highly specific for detection of poly-Neu5Ac glycotype and do not display reactivity towards poly-Neu5Gc. Naito-Matsui, Y., et al. (2017). J. Biol. Chem. 292(7); 2557-2570; Davies, LRL., et al. (2012). J. Biol. Chem. 287(34); 28917-28931; Sato, C., et al. (1995). J. Biol. Chem. 270(32); 18923-18928).

Splenocytes from mouse injected with forebrain region of E18 rat embryo.

Quality Control Testing

Evaluated by Western Blotting in one day old Mouse brain tissue extract.

Western Blotting Analysis: A 1:1,000 dilution of this antibody detected Poly-Neu5Ac in one day old Mouse brain tissue extract.

Tested Applications

Flow Cytometry Analysis: A representative lot detected Poly-Neu5Ac in Flow Cytometry applications (Davies, L.R.L., et al. (2012). J Biol Chem. 287(34):28917-31).

Western Blotting Analysis: A representative lot detected Neu5Ac in Western Blotting applications (Yabe, U., et al. (2003). J Biol Chem. 278(16):13875-80; Naito-Matsui, Y., et al. (2017). J Biol Chem. 292(7):2557-2570).

Immunoprecipitation Analysis: A representative lot detected Neu5Ac in Immunoprecipitation applications (Yabe, U., et al. (2003). J Biol Chem. 278(16):13875-80).

ELISA Analysis: A representative lot detected Neu5Ac in ELISA applications (Sato, C., et al. (1995). J Biol Chem. 270(32):18923-8).

Note: Actual optimal working dilutions must be determined by end user as specimens, and experimental conditions may vary with the end user.

Clone 12E3 is a mouse monoclonal antibody that detects poly-N-Acetylneuraminic acid (Neu5Ac).

Purified mouse monoclonal antibody IgM in PBS without preservatives.

1.0 mg/mL. Please refer to guidance on suggested starting dilutions and/or titers per application and sample type.

Store at -10°C to -25°C. Handling Recommendations: Upon receipt and prior to removing the cap, centrifuge the vial and gently mix the solution. Aliquot into microcentrifuge tubes and store at -20°C. Avoid repeated freeze/thaw cycles, which may damage IgG and affect product performance.

Concentration: Please refer to the Certificate of Analysis for the lot-specific concentration.

Unless otherwise stated in our catalog or other company documentation accompanying the product(s), our products are intended for research use only and are not to be used for any other purpose, which includes but is not limited to, unauthorized commercial uses, in vitro diagnostic uses, ex vivo or in vivo therapeutic uses or any type of consumption or application to humans or animals.