Detection of Human Glucosaminyl (N‑acetyl) Transferase 1/GCNT1 by Western Blot.
Western blot shows lysates of SW480 human colorectal adenocarcinoma cell line, TT human medullary thyroid cancer cell line, and Capan‑1 human pancreatic adenocarcinoma cell line. PVDF membrane was probed with 1 µg/mL of Sheep Anti-Human Glucosaminyl (N‑acetyl) Transferase 1/GCNT1 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF7248) followed by HRP-conjugated Anti-Sheep IgG Secondary Antibody (Catalog # HAF016). A specific band was detected for Glucosaminyl (N-acetyl) Transferase 1/GCNT1 at approximately 50 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.
Glucosaminyl (N-acetyl) Transferase 1/GCNT1 in HT‑29 human cell line.
Glucosaminyl (N-acetyl) Transferase 1/GCNT1 was detected in immersion fixed HT‑29 human colon adenocarcinoma cell line using Sheep Anti-Human Glucosaminyl (N-acetyl) Transferase 1/GCNT1 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF7248) at 15 µg/mL for 3 hours at room temperature. Cells were stained using the NorthernLights™ 557-conjugated Anti-Sheep IgG Secondary Antibody (red; Catalog # NL010) and counterstained with DAPI (blue). Specific staining was localized to plasma membrane and cytoplasm. View our protocol for Fluorescent ICC Staining of Cells on Coverslips.
Preparation and Storage
Sterile PBS to a final concentration of 0.2 mg/mL.
Reconstitution Buffer Available
The product is shipped at ambient temperature. Upon receipt, store it immediately at the temperature recommended below. *Small pack size (SP) is shipped with polar packs. Upon receipt, store it immediately at -20 to -70 °C
Stability & 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.
Mucin-type O-glycans are initiated with an O-GalNAc attachment to a serine or threonine in a polypeptide. The O-GalNAc residues are subsequently extended by various glycosyltransferases resulting in different types of O-glycans. Most O-glycans contain the core 1 structure, Gal beta 1-3GalNAc. Glucosaminyl (N-acetyl) Transferase 1 (GCNT1) converts the core 1 O-glycan to core 2 O-glycan, Gal beta 1-3(GlcNAc beta 1-6)GalNAc, via the addition of a GlcNAc residue (1, 2). Various ligand carbohydrates can be formed from core 2 branched oligosaccharides. For example, sialyl Lex in mucin‑type glycoproteins of blood cells can be formed from core 2 branched oligosaccharides (3, 4). The expression of GCNT1 was found to be associated with the progression of various types of cancer (5, 6, 7). The enzymatic activity of the recombinant GCNT1 is measured using a phosphatase-coupled method (8).
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Hatakyama, S. et al. (2010) Int. J. Cancer 127:1052.
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