Human Polypeptide GalNAc Transferase 3/GALNT3 Antibody

R&D Systems, part of Bio-Techne | Catalog # AF7174

R&D Systems, part of Bio-Techne
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AF7174
AF7174-SP
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Key Product Details

Species Reactivity

Validated:

Human

Cited:

Human

Applications

Validated:

Immunocytochemistry, Western Blot

Cited:

Mass Spectrometry, Western Blot

Label

Unconjugated

Antibody Source

Polyclonal Sheep IgG

View all Polypeptide GalNAc Transferase 3/GALNT3 Antibodies »

Product Specifications

Immunogen

Mouse myeloma cell line NS0-derived recombinant human Polypeptide GalNac Transferase 3/GALNT3
Gln38-Asp633
Accession # Q14435

Specificity

Detects human Polypeptide GalNac Transferase 3/GALNT3 in direct ELISAs and Western blots. In direct ELISAs, less than 1% cross-reactivity with recombinant human (rh) GALNT1 and rhGALNT4 is observed.

Clonality

Polyclonal

Host

Sheep

Isotype

IgG

Scientific Data Images for Human Polypeptide GalNAc Transferase 3/GALNT3 Antibody

Detection of Human and Mouse Polypeptide GalNac Transferase 3/GALNT3 antibody by Western Blot.

Detection of Human and Mouse Polypeptide GalNac Transferase 3/GALNT3 by Western Blot.

Western blot shows lysates of COLO 205 human colorectal adenocarcinoma cell line, MCF-7 human breast cancer cell line, and mouse testis tissue. PVDF membrane was probed with 0.5 µg/mL of Sheep Anti-Human Polypeptide GalNac Transferase 3/GALNT3 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF7174) followed by HRP-conjugated Anti-Sheep IgG Secondary Antibody (Catalog # HAF016). A specific band was detected for Polypeptide GalNac Transferase 3/GALNT3 at approximately 75 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.
Polypeptide GalNac Transferase 3/GALNT3 antibody in HeLa Human Cell Line by Immunocytochemistry (ICC).

Polypeptide GalNac Transferase 3/GALNT3 in HeLa Human Cell Line.

Polypeptide GalNac Transferase 3/GALNT3 was detected in immersion fixed HeLa human cervical epithelial carcinoma cell line using Sheep Anti-Human Polypeptide GalNac Transferase 3/GALNT3 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF7174) 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 Golgi granules. View our protocol for Fluorescent ICC Staining of Cells on Coverslips.

Applications for Human Polypeptide GalNAc Transferase 3/GALNT3 Antibody

Application
Recommended Usage

Immunocytochemistry

5-15 µg/mL
Sample: Immersion fixed HeLa human cervical epithelial carcinoma cell line

Western Blot

0.5 µg/mL
Sample: COLO 205 human colorectal adenocarcinoma cell line, MCF‑7 human breast cancer cell line, and mouse testis tissue

Formulation, Preparation, and Storage

Purification

Antigen Affinity-purified

Reconstitution

Sterile PBS to a final concentration of 0.2 mg/mL. For liquid material, refer to CoA for concentration.

Reconstitution Buffer Available:
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Qty

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.

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: Polypeptide GalNAc Transferase 3/GALNT3

O-glycosylation is a ubiquitous post-translational modification present in secreted and membrane‑bound proteins. Polypeptide N‑acetylgalactosaminyltransferases (GALNTs) calalyze the initial step for O-glycosylation by transferring GalNAc to Thr or Ser residues (GalNAc alpha1-O-Ser/Thr) in the Golgi compartment. Structurally, the GALNTs consist of an N-terminal catalytic domain tethered by a short linker to a C-terminal ricin-like lectin domain containing three potential carbohydrate-binding sites (1, 2). Twenty distinct GALNT isoforms have been detected in humans. These isoforms display both unique and overlapping substrate specificities (3, 4, 5) with no known universal consensus glycosylation sequence. Glycosylation of mucins results from the successive, often hierarchical, action of several specific GALNTs (6). Expression of GALNT3 appears to be highly regulated and mainly found in pancreas and testis (7). Using a peptide library screening approach, GALNT3 was classified as an intermediate transferase that increases the density of O-linked glycans within the mucin domain following glycosylation with early transferases (5). The enzymatic activity of recombinant human GALNT3 was determined using a phosphatase‑coupled assay (8).

References

  1. Gerken, T.A. et al. (2011) J. Biol. Chem. 286:14493.
  2. Ten Hagen, K.G. et al. (2003) Glycobiology 13:1R.
  3. Hagen, F.K. et al. (1997) J. Biol. Chem. 272:13843.
  4. Gerken, T.A. et al. (2006) J. Biol. Chem. 281:32403.
  5. Wandall, H.H. et al. (1997) J. Biol. Chem. 272:23503.
  6. Pratt, M.R. et al. (2004) Chem. Biol. 11:1009.
  7. Bennett, E.P. et al. (1996) J. Biol. Chem. 271:17006.
  8. Wu, Z.L. et al. (2011) Glycobiology 21:727.

Long Name

UDP-GalNAc:polypeptide N-acetylgalactosaminyltransferase 3

Alternate Names

GalNAc-T3, HFTC, HHS, pp-GaNTase 3

Entrez Gene IDs

2591 (Human); 144251 (Mouse); 366061 (Rat)

Gene Symbol

GALNT3

UniProt

Q14435 (Human)

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Product Specific Notices for Human Polypeptide GalNAc Transferase 3/GALNT3 Antibody

For research use only

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