The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
Use at an assay dependent concentration. PubMed: 24247246
Use a concentration of 1 µg/ml. Predicted molecular weight: 36 kDa. Good results were obtained when blocked with 5% non-fat dry milk in 0.05% PBS-T.
Sulfotransferase that utilizes 3'-phospho-5'-adenylyl sulfate (PAPS) to catalyze the transfer of a sulfo group to position 3 of glucosamine residues in heparan. Catalyzes the rate limiting step in the biosynthesis of heparan sulfate (HSact). This modification is a crucial step in the biosynthesis of anticoagulant heparan sulfate as it completes the structure of the antithrombin pentasaccharide binding site.
Highly expressed in the brain and kidney and weakly expressed in the heart, lung and placenta.
Anti-HS3ST1 antibody (ab91065) at 1 µg/ml (in 5% skim milk / PBS buffer) + Human fetal stomach lysate at 10 µg
Secondary HRP conjugated anti-Rabbit IgG at 1/50000 dilution
Predicted band size : 36 kDa Gel concentration: 12%
Immunocytochemistry/ Immunofluorescence - Anti-HS3ST1 antibody (ab91065)Image from Datta P et al., J Biol Chem. 2013;288(52):37308-18. Fig 3.; doi: 10.1074/jbc.M113.519033.
Cells were fixed with 4% paraformaldehyde for 15 minutes at room temperature and permeabilized with permeabilization buffer + 10% FBS in PBS for 10 minutes. Cells were incubated with ab91065 and goat anti-GM130 Golgi marker (ab1299) overnight at 4°C in the dark. Cells were washed once and stained with Alexa Fluor 647-conjugated donkey anti-rabbit IgG and Alexa Fluor 488-conjugated donkey anti-goat IgG secondary antibodies. DAPI - DNA. Dual-29 cells were used as a positive control, and the other cell samples were then evaluated at the same frequency. Overexpression of HS3st1 in Dual-29 cells resulted in a broad distribution of HS3st1. Overexpression of HS3st1 in Dual-10 cells localized HS3st1 in the Golgi. Overexpression of Golgi-targeted HS3st1 in CHO-gt31 localized HS3st1 in the Golgi.
Datta P et al. Bioengineered Chinese hamster ovary cells with Golgi-targeted 3-O-sulfotransferase-1 biosynthesize heparan sulfate with an antithrombin-binding site. J Biol Chem288:37308-18 (2013).
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