- Other Names
- TPT, HHG1, HLP3, HPE3, SMMCI, TPTPS, MCOPCB5
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- Product Citations
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Mouse Sonic Hedgehog (Shh) encodes a 437 amino acid precursor protein. It undergoes autocatalytic internal cleavage to yield a non-glycosylated 19 kD N-terminal fragment, which has signaling activity, and a glycosylated 25 kD C-terminal protein that is active in precursor processing. The Shh N-terminal fragment is highly conserved in species such as human, mouse, and rat with a greater 98% amino acids identity. Lipid modification of Shh N-terminal fragment enhances its binding affinity and modulates interactions with the Patched and Smoothened transmembrane receptors. SHH activity is attenuated by its interaction with membrane-associated Hip. On the other hand, interaction with glycans can lead to the stimulation of Shh signaling. Shh is expressed in embryonic tissues that are critical for the patterning of early embryo. It functions as the key inductive signal in the patterning of the ventral neural tube, the anterior-posterior limb axis, and the ventral somites. Shh regulates the differentiation and proliferation of early thymocyte progenitors as well as neural and hematopoietic stem cell fate. In adult tissues, Shh is associated with cancer development and tissue remodeling following an injury. Gastric Shh acts as a macrophage chemoattractant during an immune response to Helicobacter pylori. It has been shown cystein 24 replacement with two isoleucine residues confered increased activity of human Shh. Similarly, cystein 25's replacement with two isoleucines in mouse Shh resulted in increased activity of the Shh protein.Product Details
- Mouse Sonic Hedgehog (Shh), amino acids (Cys25/(Ile-Ile)-Gly198) (Accession# Q62226), with a N-terminal Met expressed in E. coli.
- Molecular Mass
- The 176 amino acid recombinant protein has a predicted molecular mass of approximately 19828.3 Da. LC-Mass Spectral data yielded a value of 19828 Da indicating the N-terminal amino acid is Met. The protein migrates at approximately 22 kD in both DTT-reducing conditions and non-reducing conditions by SDS-PAGE.
- >95%, as determined by Coomassie stained SDS-PAGE.
- 0.22 µm filtered protein solution is in PBS, pH 7.2.
- Endotoxin Level
- Less than 0.01 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 = 0.2 - 1.0 µg/mL as measured by its ability to induce C3H10T1/2 cell differentiation in the presence of hBMP9 (20 ng/mL).
- 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.
- Notochord, fetal intestine, liver, lung, and the kidney.
- SHH is essential in the patterning of early embryo, differentiation of the gastric epithelium, and T cell activation and development.
- Ventral neural tube, ventral somites, limb axis in developing embryo, and hematopoietic stem cells.
- Patched-1 (PCTH1), patched 2 (PCTH2), and smoothened (SMO).
- Mouse SHH (Cys25/(Ile-Ile)-Gly198) induces differentiation of C3H10T1/2 cells.
- Cell Type
- Neural Stem Cells, Mesenchymal Stem Cells, Hematopoietic stem and progenitors, Embryonic Stem Cells
- Biology Area
- Cell Biology, Neuroscience, Synaptic Biology, Stem Cells
- Molecular Family
- Growth Factors, Cytokines/Chemokines
- Antigen References
1. Echelard Y, et al. 1993. Cell 75:1417-1430.
2. Marigo V, et al. 1995. Genomics 28:44-51.
3. Porter JA, et al. 1996. Science. 274:255-9.
4. Zeng X, et al. 2001. Nature 411:716-20.
5. Johnson RL, Tabin CJ. 1997. Cell 90:979-90.
6. Briscoe J, Thérond PP. 2013. Nat. Rev. Mol. Cell Biol. 14:416-29.
7. Filmus J, Capurro M. 2014. Matrix Biol. 35:248-52.
8. Chuang PT, McMahon AP. 1999. Nature 397:617-21.
9. Crompton T, et al. 2007. Nat. Rev. Immunol. 7:726-35.
10. Shiotani A, et al. 2005. Am. J. Gastroenterol. 100:581-7.
11. Pepinsky RB, et al. 2000. J. Biol. Chem. 275:10995-11001.
- Gene ID
- 20423 View all products for this Gene ID
- View information about Sonic Hedgehog 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.