Recombinant Human/Mouse Indian Hedgehog (C28II) N-Term, CF

• Reactivity: Hu, Mu
• Applications: Bioactivity
• Format: Carrier-Free

Recombinant Human/Mouse Indian Hedgehog (C28II) N-Term, CF Summary

• Details of Functionality: Measured by its ability to induce alkaline phosphatase production by C3H10T1/2 mouse embryonic fibroblast cells. Nakamura, T. et al. (1997) Biochem. Biophys. Res. Commun. 237:465. The ED₅₀ for this effect is 3-12 µg/mL.
• Source: E. coli-derived Indian Hedgehog/Ihh protein
  Cys28-Gly202 (Cys28Ile-Ile), with an N-terminal Met
• Accession #: P97812.2
• N-terminal Sequence: Met
• Protein/Peptide Type: Recombinant Proteins
• Gene: IHH
• Purity: >97%, by SDS-PAGE under reducing conditions and visualized by silver stain.
• Endotoxin Note: <0.10 EU per 1 μg of the protein by the LAL method.

Applications/Dilutions

• Theoretical MW: 20 kDa.
  Disclaimer note: The observed molecular weight of the protein may vary from the listed predicted molecular weight due to post translational modifications, post translation cleavages, relative charges, and other experimental factors.

Packaging, Storage & Formulations

• 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.
  • 3 months, -20 to -70 °C under sterile conditions after reconstitution.
• Buffer: Lyophilized from a 0.2 μm filtered solution in NaH₂PO₄, NaCl and DTT with Trehalose.
• Purity: >97%, by SDS-PAGE under reducing conditions and visualized by silver stain.
• Reconstitution Instructions: Reconstitute at 100 μg/mL in sterile water.

Notes

This product is produced by and ships from R&D Systems, Inc., a Bio-Techne brand.

Alternate Names for Recombinant Human/Mouse Indian Hedgehog (C28II) N-Term, CF

• BDA1
• BDA1Indian hedgehog homolog (Drosophila)
• HHG2
• HHG-2
• Ihh
• Indian hedgehog (Drosophila) homolog
• Indian hedgehog homolog
• indian hedgehog protein
• Indian Hedgehog

Background

The hedgehog (hh) gene encoding a secreted protein was originally identified in Drosophila as a segment polarity gene. The vertebrate homologues of Hh comprise several proteins including sonic hedgehog (Shh), Indian hedgehog (Ihh), and Desert hedgehog (Dhh) (1). Hedgehog proteins are important signaling molecules during embryonic development and are highly conserved within and across species (1). Mouse and human Ihh share 100% amino acid identity in the signaling domain, while mouse Ihh and Shh share 90% amino acid identity in the N-terminal signaling domain. Ihh mRNA expression is detected in fetal lung, gut, stomach, liver, kidney, pancreas and strongly in cartilage - in growth regions of the developing bone (2, 3). Ihh, along with parathyroid hormone related protein, regulate the rate of chondrocyte proliferation and differentiation (4). Ihh is also involved in yolk sac vasculogenesis, playing an important role in differentiation of epiblast cells into endothelial and red blood cells (5).

Mouse Ihh cDNA encodes a 411 amino acid (aa) polypeptide with a predicted 27 aa signal peptide. This polypeptide is cleaved to generate a 45 kDa precursor protein that undergoes the same post‑translation processing as Shh (3). An autocatalytic reaction yields a 19 kDa amino‑terminal domain Ihh‑N protein that retains all known signaling capabilities, and a 23 kDa carboxy‑terminal domain Ihh-C protein (3). Since hydrophobic modifications to Shh, including the substitution of the N-terminal cysteine residue with two hydrophobic isoleucine residues, can also increase its potency (6), a similar modification was made for Ihh. This modified form also shows increased potency in a bioassay measuring induction of alkaline phosphatase. At the cell surface, Hedgehog activity is mediated by a multicomponent receptor complex involving the 12‑pass transmembrane protein Patched (Ptc) which binds Hedgehogs with high affinity and Smoothened (Smo), a signaling seven transmembrane G-protein coupled receptor (1).

1. Ingham, P. and A. McMahon (2001) Genes & Dev. 15:3059.
2. Marigo, V. et al. (1995) Genomics 28:44.
3. Valentini, R.P. et al. (1997) J Biol Chem. 272:8466.
4. Vortkamp, A. et al. (1996) Science 273:613.
5. Byrd, N. et al. (2002) Development 129:361.
6. Taylor, F.R. et al. (2001) Biochemistry 40: 4359.

Publications for Indian Hedgehog/Ihh (1705-HH/CF)(6)

Publications using 1705-HH/CF

• LK Veistinen, T Mustonen, MR Hasan, M Takatalo, Y Kobayashi, DA Kesper, A Vortkamp, DP Rice Regulation of Calvarial Osteogenesis by Concomitant De-repression of GLI3 and Activation of IHH Targets Front Physiol, 2017;8(0):1036. 2017 [PMID: 29311969] (Bioassay, Mouse)
• Handorf A, Chamberlain C, Li W Endogenously produced Indian Hedgehog regulates TGFbeta-driven chondrogenesis of human bone marrow stromal/stem cells. Stem Cells Dev, 2015;24(8):995-1007. 2015 [PMID: 25519748] (Bioassay, Human)
• St John, Hillary, Bishop, Kathleen, Meyer, Mark B, Benkusky, Nancy A, Leng, Ning, Kendziorski, Christin, Bonewald, Lynda F, Pike, J Wesley The osteoblast to osteocyte transition: epigenetic changes and response to the vitamin D3 hormone. Mol Endocrinol, 2014;28(7):1150-65. 2014 [PMID: 24877565] (Bioassay, Mouse)
• Saito A, Kanemoto S, Zhang Y, Asada R, Hino K, Imaizumi K Chondrocyte proliferation regulated by secreted luminal domain of ER stress transducer BBF2H7/CREB3L2. Mol Cell, 2014;53(1):127-39. 2014 [PMID: 24332809] (Bioassay, Human)
• Wilson TR, Fridlyand J, Yan Y, Penuel E, Burton L, Chan E, Peng J, Lin E, Wang Y, Sosman J, Ribas A, Li J, Moffat J, Sutherlin DP, Koeppen H, Merchant M, Neve R, Settleman J Widespread potential for growth-factor-driven resistance to anticancer kinase inhibitors. Nature, 2012;487(7408):505-9. 2012 [PMID: 22763448] (Bioassay, Human)
• Kelly MA, Hirschi KK Signaling hierarchy regulating human endothelial cell development. Arterioscler. Thromb. Vasc. Biol., 2009;29(5):718-24. 2009 [PMID: 19213939]

Bioinformatics

• Gene Symbol: IHH
• Uniprot:
  • P97812.2()