Glucosaminyl (N-acetyl) Transferase 1/GCNT1 Antibody [Unconjugated]

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 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.

Glucosaminyl (N-acetyl) Transferase 1/GCNT1 was detected in immersion fixed paraffin-embedded sections of human bladder cancer tissue using Sheep Anti-Human Glucosaminyl (N-acetyl) Transferase 1/GCNT1 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF7248) at 3 µg/mL overnight at 4 °C. Tissue was stained using the Anti-Sheep HRP-DAB Cell & Tissue Staining Kit (brown; Catalog # CTS019) and counterstained with hematoxylin (blue). Specific staining was localized to plasma membrane in smooth muscle cells. View our protocol for Chromogenic IHC Staining of Paraffin-embedded Tissue Sections.

• Reactivity: Hu
• Applications: WB, IHC, ICC/IF
• Clonality: Polyclonal
• Host: Sheep
• Conjugate: Unconjugated
• Concentration: LYOPH

Glucosaminyl (N-acetyl) Transferase 1/GCNT1 Antibody [Unconjugated] Summary

• Immunogen: Mouse myeloma cell line NS0-derived recombinant human Glucosaminyl (N-acetyl) Transferase 1/GCNT1
  Arg33-His428
  Accession # NP_001481
• Specificity: Detects human Glucosaminyl (N-acetyl) Transferase 1/GCNT1 in direct ELISAs and Western blots.
• Source: N/A
• Isotype: IgG
• Clonality: Polyclonal
• Host: Sheep
• Gene: GCNT1
• Purity Statement: Antigen Affinity-purified

Applications/Dilutions

• Dilutions:
  • Immunocytochemistry 5-15 ug/mL
  • Immunohistochemistry 3-15 ug/mL
  • Western Blot 1 ug/mL

Packaging, Storage & Formulations

• 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.
• Buffer: Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. *Small pack size (SP) is supplied either lyophilized or as a 0.2 µm filtered solution in PBS.
• Preservative: No Preservative
• Concentration: LYOPH
• Reconstitution Instructions: Sterile PBS to a final concentration of 0.2 mg/mL.

Notes

This product is produced by and ships from R&D Systems, Inc., a Bio-Techne brand.

Alternate Names for Glucosaminyl (N-acetyl) Transferase 1/GCNT1 Antibody [Unconjugated]

• beta-1,3-galactosyl-O-glycosyl-glycoproteinbeta-1,6-N-acetylglucosaminyltransferase
• beta-1,6-N-acetylglucosaminyltransferase
• C2GNT1
• C2GNTbeta-1,3-galactosyl-O-glycosyl-glycoproteinbeta-1,6-N--acetylglucosaminyltransferase
• C2GNT-L
• core 2 beta1,6 N-acetylglucosaminyltransferase-I
• core 2 beta-1,6-N-acetylglucosaminyltransferase I
• core 2 branching enzyme
• Core 2 GNT
• Core 2-branching enzyme
• core2-GlcNAc-transferase
• EC 2.4.1.102
• G6NT
• GCNT1
• glucosaminyl (N-acetyl) transferase 1, core 2
• glucosaminyl (N-acetyl) transferase 1, core 2(beta-1,6-N-acetylglucosaminyltransferase)
• MGC126335
• MGC126336
• NACGT2
• NAGCT2

Background

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 Le^x 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).

1. Bierhuizen, M.F. (1993) Genes & Development 7:468.
2. Yeh, J.C. et al. (1999) J. Biol. Chem. 274:3215.
3. Hemmerich, S. et al. (1995) J. Biol. Chem. 270:12035.
4. Wilkins, P.P. et al. (1996) J. Biol. Chem. 270:18732.
5. Shimodaira, K. et al. (1997) Cancer Res. 57: 5201.
6. Hatakyama, S. et al. (2010) Int. J. Cancer 127:1052.
7. St Hill, C.A. et al. (2009) BMC Cancer 9:79.
8. Wu, Z.L. et al. (2011) Glycobiology 21:727.

Limitations

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Primary Antibodies are guaranteed for 1 year from date of receipt.

Bioinformatics

• Gene Symbol: GCNT1
• Uniprot:
  • NP_001481()