Biolegend, 753516, Cell-Vive™ GMP Recombinant Human Sonic Hedgehog (carrier-free), 100μg

Human Sonic Hedgehog (SHH) encodes a 462 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 between species with > 98% amino acid identity between human, mouse, and rat. Lipid modifications of the 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 glycan can lead to the stimulation of SHH signaling. SHH is expressed in embryonic tissues that are critical for the patterning of early embryos. It functions as the key inductive signal in 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 and it also regulates the neural and hematopoietic stem cell fates. In adult tissues, SHH is associated with cancer development and tissue remodeling following injuries. Gastric SHH acts as a macrophage chemoattractant during the immune response to Helicobater pylori . BioLegend Cell-Vive™ GMP Recombinant proteins are manufactured and tested in accordance with USP Chapter 1043, Ancillary Materials for Cell, Gene and Tissue-Engineered Products and Ph. Eur. Chapter 5.2.12 in a dedicated GMP facility compliant with ISO 13485:2016. Specifications and processes include:
100μg
Source: Human SHH, amino acids (Cys24-Gly197) (Cys24Ile-Ile) (Accession# NP_000184), with a N-terminal Met was expressed in E. coli.
Molecular Mass: The 176 amino acid recombinant protein has a predicted molecular mass of approximately 19.8 kD. The protein migrates at approximately 22 kD in both DTT-reducing and non-reducing conditions by SDS-PAGE. The predicted N-terminal amino acid is Met.
N-terminalSequence Analysis: Met-Ile-Ile-Gly-Pro-Gly-Arg-Gly-Phe-Gly
Purity: ≥ 95%, as determined by Coomassie stained SDS-PAGE.
Formulation: 0.1 μm filtered protein solution is in 10 mM sodium phosphate buffer, 0.3M NaCl, and in pH 7.2
Endotoxin Level: Less than or equal to 0.1 EU per μg of protein as determined by the LAL method.
Residual Host Cell Protein Content: ≤ 0.500 ng/μg by ELISA
Concentration: 25 μg & 100 μg sizes are bottled at 0.5 mg/mL
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% endotoxin-free 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 or stored at -20°C or colder for up to 3 months. Avoid repeated freeze/thaw cycles.
Activity: ED50 = 0.15 - 0.6 μg/mL as measured by the ability to induce C3H10T1/2 cell differentiation in the presence of hBMP9 (20 ng/mL) as measured by alkaline phosphatase production.
Application: Bioassay
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 [email protected].
Application References(PubMed link indicates BioLegend citation): Huang WK, et al. 2021. Cell Stem Cell. 28:1657. PubMed
Disclaimer: BioLegend Cell-Vive™ GMP Recombinant proteins are for research use only. Suitable for ex vivo cell processing. Not for injection or diagnostic or therapeutic use. Not for resale. BioLegend will not be held responsible for patent infringement or other violations that may occur with the use of our products.
Function: SHH is essential in the patterning of early embryo, differentiation of the gastric epithelium, and T cells.
Interaction: Ventral neural tube, ventral somites, limb axis in developing embryo, and hematopoietic stem cells.
Ligand/Receptor: Patched-1 (PCTH1), patched 2 (PCTH2), and smoothened (SMO).
Bioactivity: SHH enhances BMP9 induced differentiation of C3H10T1/2 cells.
Cell Sources: Notochord, fetal intestine, liver, lung, and kidney
Antigen References: Echelard Y, et al. 1993. Cell. 75:1417-1430. Marigo V, et al. 1995. Genomics. 28:44-51. Porter JA, et al. 1996. Science. 5285:255-9. Zeng X, et al. 2001. Nature. 411:716-720. Johnson RL, et al. 1997. Cell. 90:979-990. Briscoe J, Therond PP. 2013. Nat. Rev. Mol. Cell Biol. 14:416-29. Filmus J, Capurro M. 2014. Matrix Biol. 35:248-52. Chuang PT, McMahon AP. 1999. Nature. 397:617-21. Crompton T, et al. 2007. Nat. Rev. Immunol. 7:726-35. Shiotani A, et al. 2005. Am. J. Gastroenterol. 100:581-7.
Gene ID: 6469
UniProt: View information about Sonic Hedgehog on UniProt.org
Other Names: SHH, TPT, HHG1, HLP3, HPE3, SMMCI, TPTPS, MCOPCB5