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Recombinant human Shh protein

QK055

Brand: Qkine

Sonic hedgehog protein (Shh) is a member of the Hedgehog family with an essential role in embryonic development, tissue regeneration, and tumorigenesis. Shh induces the cell fate and patterning of neural progenitors in ventral domains at various levels in the forebrain, midbrain, hindbrain, and spinal cord. It has many applications in the neural stem cell field where it plays a significant role in differentiating human-induced pluripotent stem cells (iPSC) towards motor neurons and interneurons. Also, it induces the patterning of organoids and embryos in culture.

Recombinant Shh protein has a molecular weight of 19.8 kDa. This protein is animal origin-free (AOF), carrier protein-free, and tag-free to ensure its purity with exceptional lot-to-lot consistency. Sonic hedgehog protein is suitable for the culture of reproducible and high-quality neurons and organoids.

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Currency: 

Product name Catalog number Pack size Price Price (USD) Price (GBP) Price (EUR)
Recombinant human Shh protein, 25 µg QK055-0025 25 µg (select above) $ 210.00 £ 155.00 € 182.00
Recombinant human Shh protein, 50 µg QK055-0050 50 µg (select above) $ 290.00 £ 210.00 € 246.00
Recombinant human Shh protein, 100 µg QK055-0100 100 µg (select above) $ 465.00 £ 340.00 € 398.00
Recombinant human Shh protein, 500 µg QK055-0500 500 µg (select above) $ 1,650.00 £ 1,200.00 € 1,402.00
Recombinant human Shh protein, 1000 µg QK055-1000 1000 µg (select above) $ 2,600.00 £ 1,900.00 € 2,220.00

Note: prices shown do not include shipping and handling charges.

Qkine company name and logo are the property of Qkine Ltd. UK.

Alternative protein names
Hhg-1, SHH, HHG1, HLP3, HPE3, MCOPCB5, SMMCI, TPT, TPTPS, sonic hedgehog, Sonic hedgehog, ShhNC, sonic hedgehog signaling molecule
Species reactivity

human

Species protein similarity:
mouse – 99%
rat – 97%
porcine – 98%
bovine – 99%

Frequently used together with:

Summary

  • High purity sonic hedgehog protein (Uniprot: Q15465)
  • >98%, by SDS-PAGE quantitative densitometry
  • 19.8 kDa (monomer)
  • Expressed in E. coli
  • Animal origin-free (AOF) and carrier protein-free
  • Manufactured in Cambridge, UK
  • Lyophilized from PBS
  • Resuspend in water at >100 µg/ml, prepare single use aliquots, add carrier protein if desired and store frozen at -20°C (short-term) or -80°C (long-term).
Handling and Storage FAQ

Featured applications

  • Neural differentiation
  • Midbrain differentiation
  • Ventral forebrain differentiation
  • Corpus organoid generation
  • Cholinergic-like neurons (ChLNs) differentiation
  • Dopaminergic neurons differentiation

Bioactivity

Human Shh Qk055 protein bioactivity lot 104376

The bioactivity of sonic hedgehog protein (Qk055) is determined by its ability to induce alkaline phosphatase production by C3H/10T1/2 (CCL-226) cells. The EC50 for this effect is 0.21 µg/ml (10.6 nM). Cells were incubated with different concentrations of Shh for 5 days before alkaline phosphatase measurement. Data is n=2, from Qk055 batch #104376.

Purity

Human Shh Qk055 protein purity SDS-PAGE lot #104391

Human Shh (Qk055) migrates as a major band at 19.8 kDa in non-reducing (NR) conditions. We load 3 ug of  protein to ensure good visibility of protein in the gel. This allows you to see the faint Shh dimer band at 40 kDa in the non-reduced sample. Shh spontaneously forms non-covalent dimers, this is thought to increase its potency (10). Upon reduction (R), only the 19.8 kDa band is visible. No contaminating protein bands are visible. Purified recombinant protein (3 µg) was resolved using 15% w/v SDS-PAGE in reduced (+β-mercaptoethanol, R) and non-reduced (-β-mercaptoethanol, NR) conditions and stained with Coomassie Brilliant Blue R-250. Data from Qk055 batch #104391.

Further quality assays

  • Mass spectrometry: single species with expected mass
  • Endotoxin: <0.005 EU/μg protein (below level of detection)
  • Recovery from stock vial:  >95%

Protein background

Sonic Hedgehog protein (Shh) is a member of the Hedgehog family, which also includes Indian-Hedgehog (Ihh) and Desert-Hedgehog (Dhh) [1-4]. Shh is essential in embryonic development, tissue regeneration, and tumorigenesis. Shh regulates cell fate and patterning of tissues, in particular the neural tube and eventually the central nervous system [5-7]. It plays a significant role in patterning ventral forebrain fate in the forebrain region, dopaminergic neural destiny in the midbrain region, and serotonergic neural fate in the hindbrain region. Shh instructs the neural progenitors’ patterning and induces motor neurons and interneurons [5]. Shh is important in adult tissue maintenance and repair during stem cell differentiation and tissue regeneration [8,9].

In cell culture, Shh has many applications in the embryonic development and neural stem cell field. Shh is used to pattern embryos and organoids towards the neural axis [10,11]. Sonic Hedgehog signaling is crucial in deriving neural progenitors from induced pluripotent stem cells [12-14]. Further differentiation into more mature neurons using a combination of different growth factors. Dopaminergic neurons are obtained using brain-derived neurotrophic factor (BDNF), glial cell line-derived neurotrophic factor (GDNF), FGF-8a/FGF-8b [15-17], TGF-β3, and Shh [15-17]. GABAergic neurons are obtained using BDNF, GDNF, insulin-like growth factor 1 (IGF-1), and Shh [18-20].

Shh is released by various cells during embryonic development, including the notochord cells forming the neural tube. Shh encodes a 462 amino acid precursor protein, which is autocatalytically processed to yield a 19 kDa N-terminal fragment (ShhN) and a 25 kDa C-terminal protein (ShhC). Lipids modify the N-terminal fragment to produce the mature, activated signaling molecule, which is then secreted from the cell [21]. SHH C24II is the bioactive N-terminal region, encompassing Cys24-Gly197 (Cys24Ile-Ile). It can signal via canonical and non-canonical pathways. Shh binds to Patched (Ptch), a transmembrane protein that plays a crucial role in the Hedgehog signaling pathway involved in tissue patterning during embryogenesis [22].

The implications of Shh dysregulations in developmental disorders and diseases such as cancer have gained interest in treatment development. Abnormal Shh signaling has been associated with basal cell carcinoma and medulloblastoma, where research is ongoing to find potential inhibitors of the Shh pathway to treat cancer [23,24]. Inhibitors to the Shh pathway are also relevant to treating development disorders such as Gorlin syndrome, which predispose affected individuals to basal cell carcinoma [25]. Finally, the role of Sonic Hedgehog in tissue regeneration makes it a potential therapeutic strategy in regenerative medicine [26].

Background references

  1. Cohen, M. M., Jr. The hedgehog signaling network. Am. J. Med. Genet. A. 123A, 5–28 (2003). doi: 10.1002/ajmg.a.20495
  2. Bürglin, T. R. The Hedgehog protein family. Genome Biol. 9, 241 (2008). doi.org/10.1186/gb-2008-9-11-241
  3. Choy, S. W. & Cheng, S. H. Hedgehog signaling. Vitam. Horm. 88, 1–23 (2012). doi: 10.1016/B978-0-12-394622-5.00001-8
  4. Carballo, G. B., Honorato, J. R., de Lopes, G. P. F. & Spohr, T. C. L. de S. e. A highlight on Sonic hedgehog pathway. Cell Commun. Signal. CCS 16, 11 (2018). doi: 10.1186/s12964-018-0220-7
  5. Litingtung, Y. & Chiang, C. Control of Shh activity and signaling in the neural tube. Dev. Dyn. 219, 143–154 (2000). doi.org/10.1002/1097-0177(2000)9999:9999<::AID-DVDY1050>3.0.CO;2-Q
  6. Patten, I. & Placzek*, M. The role of Sonic hedgehog in neural tube patterning. Cell. Mol. Life Sci. CMLS 57, 1695–1708 (2000). doi: 10.1007/PL00000652
  7. Mu, Y., Lee, S. W. & Gage, F. H. Signaling in adult neurogenesis. Curr. Opin. Neurobiol. 20, 416–423 (2010). doi: 10.1016/j.conb.2010.04.010
  8. Christie, K. & Turnley, A. Regulation of endogenous neural stem/progenitor cells for neural repair – factors that promote neurogenesis and gliogenesis in the normal and damaged brain. Front. Cell. Neurosci. 6, (2013). doi: 10.3389/fncel.2012.00070
  9. Meda, F. et al. Nerves, H2O2 and Shh: Three players in the game of regeneration. Semin. Cell Dev. Biol. 80, 65–73 (2018). doi: 10.1016/j.semcdb.2017.08.015
  10. Brady, M. V. & Vaccarino, F. M. Role of SHH in Patterning Human Pluripotent Cells towards Ventral Forebrain Fates. Cells 10, 914 (2021). doi: 10.3390/cells10040914
  11. Barak, M. et al. Human iPSC-Derived Neural Models for Studying Alzheimer’s Disease: from Neural Stem Cells to Cerebral Organoids. Stem Cell Rev. Rep. 18, 792–820 (2022). doi.org/10.1007/s12015-021-10254-3
  12. Bissonnette, C. J. et al. The Controlled Generation of Functional Basal Forebrain Cholinergic Neurons from Human Embryonic Stem Cells. Stem Cells 29, 802–811 (2011). doi: 10.1002/stem.626
  13. Liu, Y. et al. Medial ganglionic eminence–like cells derived from human embryonic stem cells correct learning and memory deficits. Nat. Biotechnol. 31, 440–447 (2013). doi: 10.1038/nbt.2565
  14. Duan, L. et al. Stem cell derived basal forebrain cholinergic neurons from Alzheimer’s disease patients are more susceptible to cell death. Mol. Neurodegener. 9, 3 (2014). doi: 10.1186/1750-1326-9-3
  15. Perrier, A. L. et al. Derivation of midbrain dopamine neurons from human embryonic stem cells. Proc. Natl. Acad. Sci. U. S. A. 101, 12543–12548 (2004). doi: 10.1073/pnas.0404700101
  16. Zeng, H. et al. Specification of Region-Specific Neurons Including Forebrain Glutamatergic Neurons from Human Induced Pluripotent Stem Cells. PLOS ONE 5, e11853 (2010). doi.org/10.1371/journal.pone.0011853
  17. Kriks, S. et al. Dopamine neurons derived from human ES cells efficiently engraft in animal models of Parkinson’s disease. Nature 480, 547–551 (2011). doi.org/10.1038/nature10648
  18. Lin, L., Yuan, J., Sander, B. & Golas, M. M. In Vitro Differentiation of Human Neural Progenitor Cells Into Striatal GABAergic Neurons. Stem Cells Transl. Med. 4, 775–788 (2015). doi: 10.5966/sctm.2014-0083
  19. Yang, N. et al. Generation of pure GABAergic neurons by transcription factor programming. Nat. Methods 14, 621–628 (2017). doi: 10.1038/nmeth.4291
  20. Gunhanlar, N. et al. A simplified protocol for differentiation of electrophysiologically mature neuronal networks from human induced pluripotent stem cells. Mol. Psychiatry 23, 1336–1344 (2018). doi.org/10.1038/mp.2017.56
  21. Hardy, R. Y. & Resh, M. D. Identification of N-terminal Residues of Sonic Hedgehog Important for Palmitoylation by Hedgehog Acyltransferase*. J. Biol. Chem. 287, 42881–42889 (2012). doi.org/10.1074/jbc.M112.426833
  22. Villavicencio, E. H., Walterhouse, D. O. & Iannaccone, P. M. The Sonic Hedgehog–Patched–Gli Pathway in Human Development and Disease. Am. J. Hum. Genet. 67, 1047–1054 (2000). doi: 10.1016/s0002-9297(07)62934-6
  23. Teglund, S. & Toftgård, R. Hedgehog beyond medulloblastoma and basal cell carcinoma. Biochim. Biophys. Acta BBA – Rev. Cancer 1805, 181–208 (2010). doi: 10.1016/j.bbcan.2010.01.003
  24. Samkari, A., White, J. & Packer, R. SHH inhibitors for the treatment of medulloblastoma. Expert Rev. Neurother. 15, 763–770 (2015). doi: 10.1586/14737175.2015.1052796
  25. Lo Muzio, L. Nevoid basal cell carcinoma syndrome (Gorlin syndrome). Orphanet J. Rare Dis. 3, 32 (2008). doi.org/10.1186/1750-1172-3-32
  26. Brockes, J. P. & Kumar, A. Appendage Regeneration in Adult Vertebrates and Implications for Regenerative Medicine. Science 310, 1919–1923 (2005). doi: 10.1126/science.1115200