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Human Glucosaminyl (N-acetyl) Transferase 1/GCNT1 Antibody

Catalog #: AF7248 Datasheet
Catalog # Availability Size / Price Qty
AF7248
AF7248-SP
Detection of Human Glucosaminyl (N‑acetyl) Transferase 1/GCNT1 by Western Blot.
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Human Glucosaminyl (N-acetyl) Transferase 1/GCNT1 Antibody Summary

Species Reactivity
Human
Specificity
Detects human Glucosaminyl (N-acetyl) Transferase 1/GCNT1 in direct ELISAs and Western blots.
Source
Polyclonal Sheep IgG
Purification
Antigen Affinity-purified
Immunogen
Mouse myeloma cell line NS0-derived recombinant human Glucosaminyl (N-acetyl) Transferase 1/GCNT1
Arg33-His428
Accession # NP_001481
Formulation
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.
Label
Unconjugated

Applications

Recommended Concentration
Sample
Western Blot
1 µg/mL
See below
Immunohistochemistry
3-15 µg/mL
See below
Immunocytochemistry
5-15 µg/mL
See below

Please Note: Optimal dilutions should be determined by each laboratory for each application. General Protocols are available in the Technical Information section on our website.

Scientific Data

Western Blot Detection of Human Glucosaminyl (N-acetyl) Transferase 1/GCNT1 antibody by Western Blot. View Larger

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.

Immunocytochemistry Glucosaminyl (N-acetyl) Transferase 1/GCNT1 antibody in HT-29 human cell line by Immunocytochemistry (ICC). View Larger

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.

Immunohistochemistry Glucosaminyl (N-acetyl) Transferase 1/GCNT1 antibody in Human Bladder Cancer Tissue by Immunohistochemistry (IHC-P). View Larger

Glucosaminyl (N-acetyl) Transferase 1/GCNT1 in Human Bladder Cancer Tissue. 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.

Preparation and Storage

Reconstitution
Sterile PBS to a final concentration of 0.2 mg/mL.
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Shipping
Lyophilized product is shipped at ambient temperature. Liquid small pack size (-SP) is shipped with polar packs. Upon receipt, store immediately at the temperature recommended below.
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.

Background: Glucosaminyl (N-acetyl) Transferase 1/GCNT1

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).

References
  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.
Long Name
Glucosaminyl (N-acetyl) Transferase 1 Core 2
Entrez Gene IDs
2650 (Human); 14537 (Mouse); 64043 (Rat)
Alternate Names
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

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