Mouse Sonic Hedgehog/Shh N-Terminus Quantikine ELISA Kit | mShh-N Qkit

• Product Details
• Product Summary
• Precision
• Recovery
• Linearity
• Additional Infomation
• Citations

Product Details

Assay Type

Solid Phase Sandwich ELISA

Format

96-well strip plate

Assay Length

4.5 hours

Sample Type & Volume Required Per Well

Cell Culture Supernates (50 µL), Serum (50 µL), Heparin Plasma (50 µL)

Sensitivity

2.37 pg/mL

Assay Range

7.81 - 500 pg/mL (Serum, Heparin Plasma, Cell Culture Supernates)

Specificity

Natural and recombinant mouse Shh-N. This kit also recognizes low levels of Shh-N in rat serum.

Interference

No significant interference observed with available related molecules.

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Product Summary

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Precision

	Intra-Assay Precision			Inter-Assay Precision
Sample	1	2	3	1	2	3
n	20	20	20	40	44	44
Mean	22.7	54.6	212	21.9	50.4	188.7
Standard Deviation	1.08	1.31	10.6	2.28	2.96	9.21
CV%	4.8	2.4	5	10.4	5.9	4.9

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Recovery

Sample Type	Average % Recovery	Range %
Serum (n=4)	98	86-109
Heparin Plasma (n=4)	97	86-108
Cell Culture Samples (n=6)	101	96-109

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Linearity

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Additional Infomation

Molecule Information

Sonic Hedgehog/Shh

Aliases

Shh

Entrez Gene IDs

6469 (Human); 20423 (Mouse)

Background

Sonic Hedgehog

Human Shh cDNA encodes a 462 amino acid (aa) residue (45 kDa) precursor protein with a 23 aa signal peptide. An autocatalytic cleavage reaction yields a 19 kDa (residues 24 - 197) amino-terminal fragment (Shh-N), and a 25 kDa (residues 198 - 462) carboxy-terminal domain (Shh-C). The N-terminal domain retains all known signaling capabilities, while the C-terminal domain is responsible for the intramolecular processing, acting as a cholesterol transferase that covalently transfers the cholesterol molecule to the C-terminus of Shh-N. When Shh is expressed in insect or mammalian cells, a palmitoyl group is also attached to the N-terminal cysteine of Shh-N via an amide linkage. Although the binding affinity to their receptors is not changed, lipid-modified Shh-N proteins are more potent than the unmodified proteins in cell-based assays. Other hydrophobic modifications to unmodified Shh-N, including the substitution of the N-terminal cysteine residue with two hydrophobic isoleucine residues, can also increase Shh-N potency. At the cell surface, Shh-N activity is mediated by a multicomponent receptor complex involving the 12-pass transmembrane protein, Patched (Ptc) which binds Shh with high affinity and Smoothened (Smo), a signaling seven transmembrane G-protein coupled receptor. In the absence of Shh, Ptc represses Smo activity. The binding of Shh to Ptc releases the basal repression of Smo by Ptc. Shh is expressed in key embryonic tissues such as the Hensen’s node, the zone of polarizing activity in the posterior limb bud, the notochord, and the floor plate of the neural tube. Shh is involved in regulating the patterning of the developing central nervous system, somite, and limb. Shh plays an important role in the development of particular tissues such as whisker, hair, foregut, tooth and bone. Studies suggest that Shh is involved in regulating stem cell fates of neural and hematopoeitic lineages, and that aberrant Shh signaling is implicated in basal cell carcinomas and other diseases.

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Citations

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