Dysregulation of cellular glycosylation is a common characteristic of many cancers (1). The Tn antigen (GalNAcα1-O-Ser/Thr), is up-regulated in a wide variety of cancers (2,3), and has been investigated as a diagnostic & prognostic marker, and as a therapeutic (4). It has also been explored as a component of anti-cancer vaccines (5). The specificity of aTn was shown by binding to asialo bovine submaxillary mucin (aBSM), which presents the Tn antigen, and not to BSM in which the Tn epitope is shielded by sialic acid residues (ref 3 and Figure 1 panel B). This antibody has previously been sold under the name HB-Tn (6,7).
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Preparation: Monoclonal antibodies were produced from mouse hybridoma 5F4 clone.
Purification: Three step liquid chromatography, proprietary.
Catalog #: SBH-Tn
Formulation: Sterile filtered solution of PBS at 0.5mg/ml.
Stability: Stable for 4 weeks at 4C. Stable for 6 months at -80C. Avoid repeated freeze-thaw cycles.
Purity: Greater than 90% by SDS-PAGE.
Specificity Assay: To create asialo bovine submaxillary mucin (aBSM), 30mg of BSM (Sigma M3895) was treated with 5U of Neuraminidase (Sigma N2876) for 2 hrs at 37⁰C. Double dilution ELISAs were carried out by coating 96 well plates with a 1:2 serial dilution of 10ng/ml BSM or aBSM, blocking and incubating with a 1:4 serial dilution of anti-Tn (SBH-Tn). After washing, plates were incubated with a goat-anti-mouse (IgG+IgM)-HRP conjugate (Thermo 31446) at 1:1000 dilution. After washing, plates were developed with 2x ABTS (Southern Biotech 0202-01). Net OD was measured at 405 – 650 nm. SBH-Tn MAb diluted 4,000 fold could detect aBSM coated at 25 ng/ml, with 30 fold less signal for BSM (see Figure 1 panel (B) above).
Usage: For research only. Not for use in diagnostic or therapeutic procedures.
Country of Origin: USA
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