Recombinant human Shh protein (Qk055)

Name: Recombinant human Shh protein (Qk055)
SKU: Qk055
Availability: InStock

Sonic hedgehog (Shh) protein has many applications in the neural stem cell field. Shh is used as a growth factor (cytokine) to induce cell types in ventral domains at various levels in the forebrain, midbrain, hindbrain, and spinal cord. It plays a significant role in patterning human-induced pluripotent stem cells (iPSC) towards ventral forebrain fate in the forebrain region, dopaminergic neuronal fate in the midbrain region, and serotonergic neuronal fate in the hindbrain region. Shh signaling is crucial in deriving motor neurons from iPSCs to model neuromuscular disorders.

SHH C24II is the bioactive N-terminal region, encompassing Cys24-Gly197 (Cys24Ile-Ile), animal-origin free (AOF) and carrier protein-free.

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1mg may be despatched as 2 x 500µg

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Description

Summary

High purity Shh (Uniprot: Q15465)
19.8 kDa (monomer)
Expressed in E. coli
Animal-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 ^oC (short-term) or -80 ^oC (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

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 – 99.4%
rat – 97.7%
porcine – 98.85%
bovine – 99.4%

Frequently used together

Recombinant Human Noggin protein
Recombinant human BDNF protein
Recombinant human GDNF protein

Bioactivity

The bioactivity of Shh (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) 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%

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We are a company founded and run by scientists to provide a service and support innovation in stem cell biology and regenerative medicine. All our products are exceptionally high purity, with complete characterisation and bioactivity analysis on every lot.

Protein background

Sonic hedgehog (Shh) protein has many applications in the neural stem cell field. It is used to induce cell types in ventral domains at various levels in the forebrain, midbrain, hindbrain, and spinal cord. It plays a significant role in patterning human-induced pluripotent stem cells (iPSC) towards ventral forebrain fate in the forebrain region, dopaminergic neuronal fate in the midbrain region, and serotonergic neuronal fate in the hindbrain region. Shh signaling is crucial in deriving motor neurons from iPSCs to model neuromuscular disorders.

Shh is a member of the Hedgehog family with an essential role in vertebrate embryonic development, patterning of the central nervous system, and tumorigenesis [1]. Shh can signal via canonical and non-canonical pathways. Human Shh encodes a 462 amino acid (aa) precursor protein which is autocatalytically processed to yield a 19 kDa N-terminal fragment (ShhN) and a 25 kDa C-terminal protein (ShhC). The N-terminal fragment is modified by lipids to produce the mature, activate signaling molecule, which is then secreted from the cell [3].

SHH C24II is the bioactive N-terminal region, encompassing Cys24-Gly197 (Cys24Ile-Ile).

Background references

1. Carballo, G.B., Honorato, J.R., de Lopes, G.P. et al. (2018) A highlight on Sonic hedgehog pathway. Cell Commun Signal 16, 11.
2. McComish SF, Caldwell MA. (2018) Generation of defined neural populations from pluripotent stem cells. Philos Trans R Soc Lond B Biol Sci. 373(1750).
3. Hardy, R. Y. & Resh, M. D. (2012) Identification of N-terminal Residues of Sonic Hedgehog Important for Palmitoylation by Hedgehog Acyltransferase. J Biol Chem 287, 42881–42889.
4. Brady, M. V. & Vaccarino, F. M. (2021) Role of SHH in Patterning Human Pluripotent Cells towards Ventral Forebrain Fates. Cells 10, 914.
5. Noguchi, Ta., Ninomiya, N., Sekine, M. et al. (2015) Generation of stomach tissue from mouse embryonic stem cells. Nat Cell Biol 17, 984–993.
6. Chakrabarti, J., Dua-Awereh, M., Schumacher, M. et al. (2022) Sonic Hedgehog acts as a macrophage chemoattractant during regeneration of the gastric epithelium. npj Regen Med 7, 3
7. Wu, S.M., Choo, A.B., Yap, M.G. and Chan, K.K.K. (2010) Role of Sonic hedgehog signaling and the expression of its components in human embryonic stem cells. Stem cell research, 4(1), pp.38-49.
8. Ho KS & Scott MP. (2002) Sonic hedgehog in the nervous system: functions, modifications and mechanisms. Curr Opin Neurobiol 12(1): 57–63.
9. Martí E. (2002) Sonic hedgehog in CNS development: one signal, multiple outputs. Trends Neurosci 25(2): 89–96.
10. Derrick DJA, Wolton K, Currie RA, Tindall MJ (2021) A mathematical model of the role of aggregation in sonic hedgehog signalling. PLoS Comput Biol 17(2): e1008562

Our products are for research use only and not for diagnostic or therapeutic use. Products are not for resale.

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