Human/Mouse Heparan Sulfate 3‑O‑Sulfotransferase 4/HS3ST4 Antibody

Catalog #: AF6085 Datasheet / COA / SDS

Catalog #	Availability	Size / Price	Qty
AF6085
AF6085-SP

Detection of Human Sulfate 3-O-Sulfotransferase 4 /HS3ST4 by Western Blot.

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Human/Mouse Heparan Sulfate 3‑O‑Sulfotransferase 4/HS3ST4 Antibody Summary

Species Reactivity

Human, Mouse

Specificity

Detects human and mouse Heparan Sulfate 3-O-Sulfotransferase4/HS3ST4 in direct ELISAs and Western blots.

Source

Polyclonal Sheep IgG

Purification

Antigen Affinity-purified

Immunogen

E. coli-derived recombinant human Heparan Sulfate 3-O-Sulfotransferase4/HS3ST4
Gly184-Lys456
Accession # Q9Y661

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

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

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Detection of Human Sulfate 3-O-Sulfotransferase 4 /HS3ST4 by Western Blot. Western blot shows lysates of U-118-MG human glioblastoma/astrocytoma cell line, Neuro-2A mouse neuroblastoma cell line, and SH-SY5Y human neuroblastoma cell line. PVDF Membrane was probed with 1 µg/mL of Sheep Anti-Human Sulfate 3-O-Sulfotransferase 4/HS3ST4 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF6085) followed by HRP-conjugated Anti-Sheep IgG Secondary Antibody (Catalog # HAF016). A specific band was detected for Sulfate 3-O-Sulfotransferase 4/HS3ST4 at approximately 58 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 8.

Immunohistochemistry

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Heparan Sulfate 3-O-Sulfotransferase 4/HS3ST4 in Human Brain. Heparan Sulfate 3-O-Sulfotransferase 4/HS3ST4 was detected in immersion fixed paraffin-embedded sections of human brain (cerebellum) using Sheep Anti-Human/Mouse Heparan Sulfate 3-O-Sulfotransferase 4/HS3ST4 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF6085) at 3 µg/mL overnight at 4 °C. Before incubation with the primary antibody, tissue was subjected to heat-induced epitope retrieval using Antigen Retrieval Reagent-Basic (Catalog # CTS013). 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 Purkinje neurons. View our protocol for Chromogenic IHC Staining of Paraffin-embedded Tissue Sections.

Reconstitution Calculator

Preparation and Storage

Reconstitution

Reconstitute at 0.2 mg/mL in sterile 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: Heparan Sulfate 3-O-Sulfotransferase 4/HS3ST4

Heparan Sulfate is a highly sulfated polysaccharide that can be found on the cell surface and within the extracellular matrix. It is typically covalently attached to the protein core of proteoglycans, such as syndecans and glypicans. Heparin, on the other hand, can be considered as a highly sulfated version of Heparan Sulfate that is predominantly found in mast cells. Both heparin and Heparan Sulfate contain disaccharide repeats of uronic acid and N‑acetylglucosamine and are modified by the same sulfotransferases (1, 2). The uronic acid residues can be sulfated at the 2-O position by Heparan Sulfate 2-O sulfotransferase (HS2ST). The N‑acetylglucosamine residues can be sulfated at the N, 3-O, and 6-O positions by N‑deacetylase/N‑sulfotransferases (NDSTs), Heparan Sulfate 3-O sulfotransferases (HS3STs) and Heparan Sulfate 6-O sulfotransferases (HS6STs) respectively. There are seven HS3STs in the human genome (3, 4). HS3ST4 and HS3ST2 are brain specific and may participate in HS-dependent neurobiologic events (5). HS3ST4 can generate tetrasulfated Heparan Sulfate disaccharide, the most highly sulfated sugar found in biological samples (6, 7), and may have a role in assisting HSV-1 entry and spread (8). HS3ST4 is a Golgi resident type II membrane protein and has the longest proline rich stem region among all HS3STs (3, 5). The enzyme activity was assayed using an SDS-PAGE based method (9).

References

Bernfield, M. et al. (1999) Annu. Rev. Biochem. 68:729.
Esko, J.D. and Selleck, S.B. (2002) Annu. Rev. Biochem. 71:435.
Shworak, N.W. et al. (1999) J. Biol. Chem. 274:5170.
Xu, D. et al. (2005) Biochem. J. 386:451.
Lawrence, R. et al. (2007) Matrix Biol. 26:442.
Mochizuki, H. et al. (2003) J. Biol. Chem. 278:26780.
Wu, Z.L. et al. (2004) J. Biol. Chem. 279:1861.
Tiwari,V. et al. (2005) Biochem. Biophys. Res. Commun. 338:930.
Wu, Z.L. et al. (2010) BMC Biotechnol. 10:11.

Entrez Gene IDs

9951 (Human)

Alternate Names

30ST4; 3OST4 heparan sulfate 3-O-sulfotransferase-4; 3-OST-4; EC 2.8.2; EC 2.8.2.23; h3-OST-4; heparan sulfate (glucosamine) 3-O-sulfotransferase 4; Heparan sulfate 3-O-sulfotransferase 4; Heparan sulfate D-glucosaminyl 3-O-sulfotransferase 4; heparan sulfate glucosamine 3-O-sulfotransferase 4; HS3ST4