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The hedgehog family of morphogens are regulators of cell proliferation, differentiation, and cell-cell communication. They have important roles in organogenesis, spermatogenesis, stem cell maintenance, and oncogenesis. The vertebrate hedgehog family comprises of sonic hedgehog (Shh), Indian hedgehog (IHH or Ihh), and the Desert hedgehog (Dhh). Hedgehog proteins are highly conserved within and across species with mouse and human IHH sharing 100% amino acid identity in the signaling domain, while mouse IHH and Shh share 90% amino acid identity in the N-terminal signaling domain. The C-terminal domain displays an autoproteolysis activity and a cholesterol transferase activity. Both activities result in the cleavage of the full-length protein and covalent attachment of a cholesterol moiety to the C-terminal of the newly generated N-terminal signaling domain. Hedgehogs bind to the patched (PTC) receptor, which functions in the association with smoothened (SMO), to activate the transcription of target genes. In the embryo, IHH is detected in the developing gut, the growth zone of cartilage of developing long bones, epithelium, and the urogenital sinus. IHH is implicated in the endochondral ossification by regulating the balance between the growth and ossification of the developing bones. IHH is also known to induce the expression of parathyroid hormone-related protein (PTHRP). Since it has been shown that the replacement of N-terminal cysteine with two isoleucine residues conferred increased activity of Shh, similar modification was made for IHH.Product Details
- Mouse IHH, amino acids (Cys28/(Ile-Ile)-Gly202) (Accession# P97812.2), was expressed in E. coli.
- Molecular Mass
- The 177 amino acid recombinant protein has a predicted molecular mass of approximately 19.9 kD. The protein migrates at approximately 24 kD in both DTT-reducing and non-reducing conditions by SDS-PAGE. The predicted N-terminal amino acid is Met.
- >95%, as determined by Coomassie stained SDS-PAGE.
- 0.22 µm filtered protein solution is in PBS, 2mM CHAPS, pH 7.2.
- Endotoxin Level
- Less than 0.1 ng per µg cytokine as determined by the LAL method.
- 10 and 25 µg sizes are bottled at 200 µg/mL. 100 µg size and larger sizes are lot-specific and bottled at the concentration indicated on the vial (please contact technical support for concentration, or use our Lookup tool if you have a lot number.) Please note, new lots of the 100 µg size will be lot-specific and may differ from previous lots that had a fixed concentration.
- Storage & Handling
- Unopened vial can be stored between 2°C and 8°C for up to 2 weeks, at -20°C for up to six months, or at -70°C or colder until the expiration date. For maximum results, quick spin vial prior to opening. The protein can be aliquoted and stored at -20°C or colder. Stock solutions can also be prepared at 50 - 100 µg/mL in appropriate sterile buffer, carrier protein such as 0.2 - 1% BSA or HSA can be added when preparing the stock solution. Aliquots can be stored between 2°C and 8°C for up to one week and stored at -20°C or colder for up to 3 months. Avoid repeated freeze/thaw cycles.
- ED50 = 1.0 - 4.0 µg/mL as measured by its ability to induce C3H10T1/2 cell differentiation in the presence of hBMP9 (20 ng/mL) as measured by alkaline phosphatase production.
- Application Notes
BioLegend carrier-free recombinant proteins provided in liquid format are shipped on blue-ice. Our comparison testing data indicates that when handled and stored as recommended, the liquid format has equal or better stability and shelf-life compared to commercially available lyophilized proteins after reconstitution. Our liquid proteins are verified in-house to maintain activity after shipping on blue ice and are backed by our 100% satisfaction guarantee. If you have any concerns, contact us at firstname.lastname@example.org.
- Additional Product Notes
View more applications data for this product in our Scientific Poster Library.
Developing gut, growth zone of cartilage, epithelium, and urogenital sinus.
- Organogenesis, spermatogenesis, stem cell maintenance, and oncogenesis.
- Chondrocytes and epiblast cells.
- Patched-1 (PCTH1), patched 2 (PCTH2), and smoothened (SMO).
- Mouse IHH (Cys28/(Ile-Ile)-Gly202) induces differentiation of C3H10T1/2 cells.
- Cell Type
- Hematopoietic stem and progenitors, Mesenchymal Stem Cells, Neural Stem Cells
- Biology Area
- Stem Cells
- Molecular Family
- Cytokines/Chemokines, Growth Factors
- Antigen References
1. Berman DM, et al. 2003. Nature 425:846-851.
2. Chung U, et al. 2001. J. Clin. Invest. 107:295-304.
3. Gao B, et al. 2009. Nature 458:1196-1200.
4. Hebrok M, et al. 2000. Development 127:4905-4913.
5. Lanske B, et al. 1996. Science 273:663-666.
6. Lee K, et al. 2006. Nature Genet. 38:1204-1209.
7. Valentini RP, et al. 1997. J. Biol. Chem. 272:8466-73.
8. Vortkamp A, et al. 1996. Science 273:613-622.
- Gene ID
- 16147 View all products for this Gene ID
- View information about IHH on UniProt.org
- Does specific activity of a recombinant protein vary between lots?
Specific activity will vary for each lot and for the type of experiment that is done to validate it, but all passed lots will have activity within the established ED50 range for the product and we guarantee that our products will have lot-to-lot consistency. Please conduct an experiment-specific validation to find the optimal ED50 for your system.
- Have your recombinants been tested for stability?
Our testing shows that the recombinant proteins are able to withstand room temperature for a week without losing activity. In addition the recombinant proteins were also found to withstand four cycles of freeze and thaw without losing activity.
- How do you convert activity as an ED50 in ng/ml to a specific activity in Units/mg?
- Use formula Specific activity (Units/mg) = 10e6/ ED50 (ng/mL)
- How does the activity of your recombinant proteins compare to competitors?
We quality control each and every lot of recombinant protein. Not only do we check its bioactivity, but we also compare it against other commercially available recombinant proteins. We make sure each recombinant protein’s activity is at least as good as or better than the competition’s. In order to provide you with the best possible product, we ensure that our testing process is rigorous and thorough. If you’re curious and eager to make the switch to BioLegend recombinants, contact your sales representative today!
- What is the specific activity or ED50 of my recombinant protein?
The specific activity range of the protein is indicated on the product datasheets. Because the exact activity values on a per unit basis can largely fluctuate depending on a number of factors, including the nature of the assay, cell density, age of cells/passage number, culture media used, and end user technique, the specific activity is best defined as a range and we guarantee the specific activity of all our lots will be within the range indicated on the datasheet. Please note this only applies to recombinants labeled for use in bioassays. ELISA standard recombinant proteins are not recommended for bioassay usage as they are not tested for these applications.