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biological source
mouse
Quality Level
antibody form
purified antibody
antibody product type
primary antibodies
clone
12E3, monoclonal
purified by
affinity chromatography
species reactivity
human, mouse
species reactivity (predicted by homology)
rat
packaging
antibody small pack of 100
technique(s)
ELISA: suitable
flow cytometry: suitable
immunoprecipitation (IP): suitable
western blot: suitable
isotype
IgMκ
epitope sequence
Unknown
Protein ID accession no.
UniProt accession no.
storage temp.
-10 to -25°C
Specificity
Clone 12E3 is a mouse monoclonal antibody that detects poly-N-Acetylneuraminic acid (Neu5Ac).
Immunogen
Splenocytes from mouse injected with forebrain region of E18 rat embryo.
Application
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.
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.
Target description
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).
Physical form
Purified mouse monoclonal antibody IgM in PBS without preservatives.
Reconstitution
1.0 mg/mL. Please refer to guidance on suggested starting dilutions and/or titers per application and sample type.
Storage and Stability
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.
Other Notes
Concentration: Please refer to the Certificate of Analysis for the lot-specific concentration.
Disclaimer
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.
WGK
WGK 2
Flash Point(F)
Not applicable
Flash Point(C)
Not applicable
Regulatory Information
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Certificates of Analysis (COA)
Search for Certificates of Analysis (COA) by entering the products Lot/Batch Number. Lot and Batch Numbers can be found on a product’s label following the words ‘Lot’ or ‘Batch’.
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