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| Overview |
SHH stands for sonic hedgehog. Human SHH (C24II) is a recombinant protein optimized for use in cell culture, differentiation studies, and functional assays.
Human SHH (C24II) can be used for the expansion or differentiation of various cell types, predominantly of human and mouse embryonic stem cells, but also of adult stem cells and mesenchymal cells and others. |
| Details |
Background information
SHH is involved in a wide variety of embryonic developmental events, including neurogenesis, limb development, hematopoiesis, and gut formation. Besides their role as morphogens, hedgehog family members have also been shown to act as mitogens, cell survival factors, and axon guidance factors and to influence tissue regeneration within the adult. SHH is synthesized as a precursor, which is autocatalytically cleaved into a 19 kDa N-terminal domain (SHH-N) and a 25 kDa C-terminal domain (SHH-C). SHH-N is responsible for the signaling activity of hedgehog proteins, while SHH-C acts as a cholesterol transferase and covalently attaches a cholesterol molecule to SHH-N. The lipid-modified SHH-N displays enhanced activity. Human SHH (C24II) corresponds to the mature SHH-N domain and carries a Cys to Ile-Ile substitution resulting in a lipophilic moiety.
Quality description
Research-grade cytokines are suitable for a wide variety of cell culture applications. They are sterile-filtered prior to lyophilization. Generally, endotoxin levels are <0.1 ng/µg (<1 EU/µg), and purities are >95%. The biological activity is tested in appropriate bioassays.
Premium-grade cytokines offer the convenience of high and well-defined biological activities and allow exact unit dosing for demanding applications. The biological activity is determined after lyophilization and reconstitution, and normalized to WHO/NIBSC standards whenever available. In general, endotoxin levels are <0.01 ng/µg (<0.1 EU/µg), and purities are >97%. Lot-specific certificates of analysis are available on request (macstec@miltenyibiotec.de). |
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| Figure 1 |
| Neural precursors, derived from mouse embryonic stem cells, were expanded in MACS Neuro Medium containing N2-supplements, laminin, Human FGF-2 (10 ng/mL), Human SHH (C24II) (200 ng/mL), and FGF-8b (100 ng/mL). Subsequently cells were grown in MACS Neuro Medium complemented by B27 supplements and ascorbic acid (200 µM) for final differentiation into neurons. Cells were analyzed by immunocytochemistry for TuJ1, TH, and GAD67 expression. |
| 1: TH (tyrosine hydroxylase)+ cells |
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| 2: TuJ1 (neuron-specific β-III tubulin)+ cells |
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| 3: GAD67 (glutamate decarboxylase)+ cells |
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| Figure 2 |
| Human SHH (C24II) activity assay. The biological activity of Human SHH (C24II) was determined by alkaline phosphatase assay using mouse C3H10T1/2 fibroblasts. |
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| Figure 3 |
| SDS-PAGE of Human SHH (C24II), premium grade under reduced (R) and non-reduced (NR) conditions. |
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