>95%, by SDS-PAGE visualized with Silver Staining and quantitative densitometry by Coomassie® Blue Staining.
<0.10 EU per 1 μg of the protein by the LAL method.
Measured by its ability to inhibit Sonic Hedgehog (Shh) induction of alkaline phosphatase production in C3H10T1/2 mouse embryonic fibroblast cells. Recombinant Human Hip (Catalog # 9280-HP) inhibits a constant dose of 20 ng/mL of Recombinant Human Sonic Hedgehog/Shh Protein, High Activity
in a C3H10T1/2 mouse embryonic fibroblast cell alkaline phosphatase induction assay with an ED50 = 0.1-0.6 µg/mL.
Recombinant Human Sonic Hedgehog/Shh (Catalog # 8908-SH) induces alkaline phosphatase production in C3H10T1/2 cells (green line). Recombinant Human Hip (Catalog # 9280-HP) inhibits a constant dose of 20 ng/mL of Recombinant Human Sonic Hedgehog/Shh Protein, High Activity (Catalog # 8908-SH) in C3H10T1/2 mouse embryonic fibroblasts with an ED50 = 0.1-0.6 µg/mL (orange line).
Hedgehog signaling proteins act as mitogens, morphogens, or inducing factors in many different cell types during embryonic development. They aid in growth, patterning, and morphogenesis in both vertebrates and insects (1). Hip (Hedgehog-interacting protein) is a type I transmembrane protein identified for its ability to bind biologically active Sonic Hedgehog. It is comprised of 700 amino acids (aa), and includes a hydrophobic signal sequence, two EGF-like domains near the C-terminus, and a 22 aa membrane-attachment helix at the C-terminal end (2, 3). Mouse and human Hip share 94% aa sequence identity through the entire protein sequence (4). Alternative splicing generates a soluble isoform that lacks the C-terminal half of the protein (5). Hip has only been identified in vertebrates and binds all three mammalian Hedgehogs: sonic (Shh), desert (Dhh), and Indian (Ihh). Like the Hedgehog receptor Patched, Hip is a transcriptional target of Hedgehog signaling (2). Unlike Patched, Hip's ability to bind hedgehogs is not involved in transducing a signal intracellularly, rather it regulates the availability of Hedgehog ligand extracellularly (6). Transgenic mice overexpressing Hip in proliferating chondrocytes display skeletal defects similar to those observed in Ihh mutant mice. These results indicate that Hip is involved in attenuating Hedgehog signaling (2). The expression pattern of Hip correlates with its ability to interact with all three mammalian Hedgehogs. It is expressed in a variety of organs, adjacent with sites of hedgehog expression. For instance, Shh is expressed in the epithelium of the lung, and Hip is found in the underlying lung mesenchyme (2). In fact, Hip knock-out mice exhibit neonatal lethality with respiratory failure due to defective branching morphogenesis. This phenotype correlates with altered expression of Shh markers suggesting an increase in Shh signaling (6). Interestingly, other developmental mechanisms that rely on normal Shh signaling, such as dorsal-ventral patterning of the neural tube, development of the somites, and organ laterality appeared histologically normal in Hip-/- mice (6). Hip is also expressed in vascular endothelial cells but is down-regulated during angiogenesis and in various tumors (7, 8).
Ramsbottom, S.A. and M.E. Pownall (2016) J. Dev. Biol. 4:23.
Chuang, P-T. and A.P. McMahon (1999) Nature 397:617.
Bishop, B. et al. (2009) Nat. Struct. Mol. Biol. 16:698.
Bak, M. et al. (2001) Cytogenet. Cell Genet. 92:300.
Coulombe, J. et al. (2004) Mol. Cell. Neurosci. 25:323.
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