Heparin biotin

Manufactured by Merck Group

Heparin-biotin is a laboratory reagent that consists of the biomolecule heparin covalently linked to biotin. It serves as a tool for researchers to study and manipulate biological systems that involve heparin-binding proteins or heparin-dependent processes.

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Lab products found in correlation

P nitrophenyl phosphate, by Merck Group (1 mentions) Streptavidin, by Thermo Fisher Scientific (1 mentions) Biotinylated bovine serum albumin, by Merck Group (1 mentions)

2 protocols using heparin biotin

Heparin-Binding Assay for Murine Gammaherpesvirus

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Streptavidin-coated 96-well plates (catalog no. 15124, Pierce) were incubated (4°C, 2 hours) with heparin-biotin (1 mg ml⁻¹, catalog no. B9806, Sigma), washed with PBS, and blocked in PBS/1% BSA. WT and gp150⁻ MuHV-4 virions (10⁵ plaque-forming units per well) were then bound in the presence of increasing amounts of soluble heparin (catalog no. H3393, Sigma) for 2 hours at 37°C. Plates were washed twice with PBS, and bound virions were detected with a mAb recognizing gN (mAb 3f7) (44 (link)). Bound antibodies were detected with alkaline phosphatase–conjugated goat anti-mouse Ig pAb (Sigma). Washing was performed with PBS. p-Nitrophenylphosphate (Sigma) was used as the substrate, and absorbance was read at 405 nm using a Benchmark ELISA plate reader (Thermo Fisher Scientific). Results were expressed as the percentage of absorbance without soluble heparin.

Delguste M., Zeippen C., Machiels B., Mast J., Gillet L, & Alsteens D. (2018). Multivalent binding of herpesvirus to living cells is tightly regulated during infection. Science Advances, 4(8), eaat1273.

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Quantifying LPL Activity in Conditioned Media and on Surfaces

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For LPL activity in conditioned media, plasmids bearing WT or mutant LPL variants were transiently transfected into HEK293 cells. LPL was released from the surface of cells through the addition of heparin, media was collected and the LPL secreted in media was assayed for activity using 10 μM DGGR essentially as previously described^{35 (link)}.
For LPL activity on a surface, a 96-well black-sided, clear-bottom plate, was functionalized by a brief incubation with biotinylated bovine serum albumin (Sigma, 1 mg/ml, 5 minutes), washed three times with PBS, incubated with streptavidin (Invitrogen, 0.1 mg/ml, 20 minutes) and washed again with PBS. Plates sat on ice with PBS in the wells until the experiment was ready to proceed. Wells were then treated as follows: A) No LPL (Buffer), B) biotinylated bovine LPL (~150 nM, 5 minute incubation), C) Heparin-Biotin (Sigma, 1 mg/ml, 5 minutes) + bovine LPL (150 nM, 5 minutes). Wells were washed three more times with PBS and substrate was added (150 mM NaCl, 0.4% Triton-X, 20 mM Tris 8.0, 14 μM DGGR). Wells were excited at 529 nm with a 590 nm cut-off and read for emission at 600 nm on a SpectaMax 5 plate reader.

Hayne C.K., Yumerefendi H., Cao L., Gauer J.W., Lafferty M.J., Kuhlman B., Erie D.A, & Neher S.B. (2018). We FRET So You Don’t Have To: New Models of the Lipoprotein Lipase Dimer. Biochemistry, 57(2), 241-254.

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