Mouse Sonic Hedgehog/Shh N-Terminus Quantikine ELISA Kit Summary
Product Summary
Precision
Cell Culture Supernates, Serum, Heparin Plasma
| Intra-Assay Precision | Inter-Assay Precision | |||||
|---|---|---|---|---|---|---|
| Sample | 1 | 2 | 3 | 1 | 2 | 3 |
| n | 20 | 20 | 20 | 40 | 44 | 44 |
| Mean (pg/mL) | 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 |
Recovery
The recovery of mouse Shh-N spiked to levels throughout the range of the assay in various matrices was evaluated.
| Sample Type | Average % Recovery | Range % |
|---|---|---|
| Cell Culture Samples (n=6) | 101 | 96-109 |
| Heparin Plasma (n=4) | 97 | 86-108 |
| Serum (n=4) | 98 | 86-109 |
Linearity
Scientific Data
Product Datasheets
Preparation and Storage
Background: Sonic Hedgehog/Shh
Shh (Sonic Hedgehog) is expressed in embryonic tissues that are critical for the patterning of the developing central nervous system, somite, and limb. It is also involved in whisker, hair, foregut, tooth, and bone development. Shh regulates neural and hematopoietic stem cell fate and is important for thymocyte differentiation and proliferation. In adult tissue Shh is associated with cancer development and tissue remodeling following injury. Palmitoylation and cholesterol addition of Shh contribute to its membrane tethering and assembly into multimers. Lipid modification and multimerization greatly increase Shh signaling potency. Shh can be released from the plasma membrane by the cooperative action of DISP1, SCUBE2, and TACE/ADAM17. Canonical signaling of Shh is mediated by a receptor complex that includes Patched (PTCH1, PTCH2) and Smoothened (SMO).
Assay Procedure
Refer to the product- Prepare all reagents, standard dilutions, and samples as directed in the product insert.
- Remove excess microplate strips from the plate frame, return them to the foil pouch containing the desiccant pack, and reseal.
- Add 50 µL of Assay Diluent to each well.
- Add 50 µL of Standard, Control, or sample to each well. Cover with a plate sealer, and incubate at room temperature for 2 hours.
- Aspirate each well and wash, repeating the process 4 times for a total of 5 washes.
- Add 100 µL of Conjugate to each well. Cover with a new plate sealer, and incubate at room temperature for 2 hours.
- Aspirate and wash 5 times.
- Add 100 µL Substrate Solution to each well. Incubate at room temperature for 30 minutes. PROTECT FROM LIGHT.
- Add 100 µL of Stop Solution to each well. Read at 450 nm within 30 minutes. Set wavelength correction to 540 nm or 570 nm.
Citations for Mouse Sonic Hedgehog/Shh N-Terminus Quantikine ELISA Kit
R&D Systems personnel manually curate a database that contains references using R&D Systems products. The data collected includes not only links to publications in PubMed, but also provides information about sample types, species, and experimental conditions.
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Citations: Showing 1 - 3
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Comparison of Differentiation Pattern and WNT/SHH Signaling in Pluripotent Stem Cells Cultured under Different Conditions
Authors: B ?wierczek-, D Dudka, D Bauer, T Czajkowski, K Ilach, W Streminska, A Kominek, K Piwocka, MA Ciemerych, K Archacka
Cells, 2021-10-14;10(10):.
Species: Mouse
Sample Types: Cell Culture Supernates
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The ESCRT-III Protein CHMP1A Mediates Secretion of Sonic Hedgehog on a Distinctive Subtype of Extracellular Vesicles
Authors: ME Coulter, CM Dorobantu, GA Lodewijk, F Delalande, S Cianferani, VS Ganesh, RS Smith, ET Lim, CS Xu, S Pang, ET Wong, HGW Lidov, ML Calicchio, E Yang, DM Gonzalez, TM Schlaeger, GH Mochida, H Hess, WA Lee, MK Lehtinen, T Kirchhause, D Haussler, FMJ Jacobs, R Gaudin, CA Walsh
Cell Rep, 2018-07-24;24(4):973-986.e8.
Species: Mouse
Sample Types: CSF
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Spatially heterogeneous choroid plexus transcriptomes encode positional identity and contribute to regional CSF production.
Authors: Lun M, Johnson M, Broadbelt K, Watanabe M, Kang Y, Chau K, Springel M, Malesz A, Sousa A, Pletikos M, Adelita T, Calicchio M, Zhang Y, Holtzman M, Lidov H, Sestan N, Steen H, Monuki E, Lehtinen M
J Neurosci, 2015-03-25;35(12):4903-16.
Species: Mouse
Sample Types: Cell Culture Supernates
FAQs
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Assay Diluent RD1-63 contains visible precipitate. Is this acceptable to use?
A small amount of precipitate is expected and acceptable in Assay Diluent RD1-63. Variability has been observed between vials and diluent lots. It may not fully resolubilize but this will not impact assay performance. It is not recommended to filter the precipitate.
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