Mouse Sonic Hedgehog/Shh C-Terminus Antibody

Catalog #: AF445
11 Citations Datasheet / COA / SDS

Catalog #	Availability	Size / Price	Qty
AF445
AF445-SP

Detection of Human and Mouse Sonic Hedgehog/Shh by Western Blot.

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Citations (11)

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Mouse Sonic Hedgehog/Shh C-Terminus Antibody Summary

Species Reactivity

Mouse

Specificity

Detects mouse Shh C-terminal peptide in direct ELISAs and Western blots. In these formats, less than 5% cross-reactivity with recombinant mouse (rm) Dhh C-terminal peptide is observed and less than 1% cross-reactivity with a 6X histidine-tagged rmShh N‑terminal peptide (amino acids 25-198) is observed. Detects human and mouse Sonic Hedgehog protein in Western blots. Shh C-terminal peptide specific IgG was purified by first passing the goat sera over a mouse Shh N-terminal peptide affinity column. The unbound fraction from the mouse Shh N-terminal peptide affinity column was subsequently purified using a mouse Shh C-terminal peptide affinity column.

Source

Polyclonal Goat IgG

Purification

Antigen Affinity-purified

Immunogen

E. coli-derived recombinant mouse Sonic Hedgehog
Cys25-Ser437 with a Lys122Arg substitution
Accession # Q62226

Formulation

Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. *Small pack size (SP) is supplied either lyophilized or as a 0.2 µm filtered solution in PBS.

Label

Unconjugated

Applications

Recommended Concentration

Sample

Western Blot

1 µg/mL

See below

Immunohistochemistry

5-15 µg/mL

See below

Please Note: Optimal dilutions should be determined by each laboratory for each application. General Protocols are available in the Technical Information section on our website.

Scientific Data

Western Blot

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Detection of Human and Mouse Sonic Hedgehog/Shh by Western Blot. Western blot shows lysates of human stomach tissue and mouse stomach tissue. PVDF membrane was probed with 1 µg/mL of Goat Anti-Mouse Sonic Hedgehog/Shh C-Terminus Antigen Affinity-purified Polyclonal Antibody (Catalog # AF445) followed by HRP-conjugated Anti-Goat IgG Secondary Antibody (Catalog # HAF109). A specific band was detected for Sonic Hedgehog/Shh at approximately 50 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.

Immunohistochemistry

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Sonic Hedgehog in Mouse Spinal Cord. Sonic Hedgehog was detected in immersion fixed frozen sections of mouse spinal cord using 15 µg/mL Mouse Sonic Hedgehog C-Terminus Antigen Affinity-purified Polyclonal Antibody (Catalog # AF445) overnight at 4 °C. Tissue was stained with the Anti-Goat HRP-DAB Cell & Tissue Staining Kit (brown; Catalog # CTS008) and counterstained with hematoxylin (blue). View our protocol for Chromogenic IHC Staining of Frozen Tissue Sections.

Reconstitution Calculator

Preparation and Storage

Reconstitution

Reconstitute at 0.2 mg/mL in sterile PBS.

Shipping

The product is shipped at ambient temperature. Upon receipt, store it immediately at the temperature recommended below. *Small pack size (SP) is shipped with polar packs. Upon receipt, store it immediately at -20 to -70 °C

Stability & Storage

Use a manual defrost freezer and avoid repeated freeze-thaw cycles.

12 months from date of receipt, -20 to -70 °C as supplied.
1 month, 2 to 8 °C under sterile conditions after reconstitution.
6 months, -20 to -70 °C under sterile conditions after reconstitution.

Background: Sonic Hedgehog/Shh

The hedgehog (hh) gene encoding a secreted protein was originally identified in Drosophila as a segment polarity gene. The vertebrate homologues of Hh comprise several proteins including sonic hedgehog (Shh), Indian hedgehog (Ihh), and Desert hedgehog (Dhh). Hedgehog proteins are important signaling molecules during embryonic development. Shh genes are highly conserved and have been identified in a variety of species including human, mouse, frog, fish, and chicken. Mouse and human Shh are 92% identical at the amino acid sequence level. 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. Evidence also suggests 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.

Mouse Shh cDNA encodes a 437 amino acid residue with a predicted 24 aa residue signal peptide that is cleaved to generate a 413 aa residue precursor protein. An autocatalytic reaction yields a 19 kDa amino-terminal domain Shh-N protein containing cholesterol and palmitate, and a 27 kDa carboxy-terminal domain Shh-C protein. 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. Shh can act as both a short-range contact dependent factor and as a long-range, diffusible morphogen. At the cell surface, Shh 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.

Entrez Gene IDs

6469 (Human); 20423 (Mouse)

Alternate Names

HHG1; HHG-1; HLP3; HPE3; MCOPCB5; MCOPCB5sonic hedgehog (Drosophila) homolog; Shh; ShhNC; SMMCI; SMMCIsonic hedgehog homolog (Drosophila); sonic hedgehog homolog; sonic hedgehog protein; Sonic Hedgehog; TPT; TPTPS

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Citations for Mouse Sonic Hedgehog/Shh C-Terminus Antibody

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.

11 Citations: Showing 1 - 10
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Hedgehog proteins create a dynamic cholesterol interface
Authors: A Mafi, R Purohit, E Vielmas, AR Lauinger, B Lam, YS Cheng, T Zhang, Y Huang, SK Kim, WA Goddard, AE Ondrus
PLoS ONE, 2021-02-25;16(2):e0246814.
Species: Human
Sample Types: Cell Lysates
Applications: Western Blot
Dual roles of the sterol recognition region in Hedgehog protein modification
Authors: Rahul Purohit, Daniel S. Peng, Erika Vielmas, Alison E. Ondrus
Communications Biology
Progenitors oppositely polarize WNT activators and inhibitors to orchestrate tissue development
Authors: Irina Matos, Amma Asare, John Levorse, Tamara Ouspenskaia, June de la Cruz-Racelis, Laura-Nadine Schuhmacher et al.
eLife
Loss of GATA4 causes ectopic pancreas in the stomach
Authors: Elisa Rodríguez‐Seguel, Laura Villamayor, Noelia Arroyo, Mónica P De Andrés, Francisco X Real, Franz Martín et al.
The Journal of Pathology
The ciliopathy gene ftm/rpgrip1l controls mouse forebrain patterning via region-specific modulation of hedgehog/gli signaling
Authors: A Andreu-Cer, I Anselme, A Karam, C Laclef, M Catala, S Schneider-
J. Neurosci., 2019-01-28;0(0):.
Species: Mouse
Sample Types: Embryo
Applications: IHC
Loss of the transcription factor GLI1 identifies a signaling network in the tumor microenvironment mediating KRAS oncogene-induced transformation.
Authors: Mills, Lisa D, Zhang, Yaqing, Marler, Ronald J, Herreros-Villanueva, Marta, Zhang, Lizhi, Almada, Luciana, Couch, Fergus, Wetmore, Cynthia, Pasca di Magliano, Marina, Fernandez-Zapico, Martin E
J Biol Chem, 2013-03-12;288(17):11786-94.
Species: Mouse
Sample Types: Whole Tissue
Applications: IHC-P
T cells enhance stem-like properties and conditional malignancy in gliomas.
Authors: Irvin DK, Jouanneau E, Duvall G
PLoS ONE, 2010-06-07;5(6):e10974.
Species: Mouse
Sample Types: Whole Cells
Applications: Flow Cytometry
GLI1 is regulated through Smoothened-independent mechanisms in neoplastic pancreatic ducts and mediates PDAC cell survival and transformation.
Authors: Nolan-Stevaux O, Lau J, Truitt ML, Chu GC, Hebrok M, Fernandez-Zapico ME, Hanahan D
Genes Dev., 2009-01-01;23(1):24-36.
Species: Mouse
Sample Types: Whole Tissue
Applications: IHC-P
Characterization of progenitor domains in the developing mouse thalamus.
Authors: Vue TY, Aaker J, Taniguchi A
J. Comp. Neurol., 2007-11-01;505(1):73-91.
Species: Mouse
Sample Types: Whole Tissue
Applications: IHC-Fr
Blockade of hedgehog signaling inhibits pancreatic cancer invasion and metastases: a new paradigm for combination therapy in solid cancers.
Authors: Feldmann G, Dhara S, Fendrich V, Bedja D, Beaty R, Mullendore M, Karikari C, Alvarez H, Iacobuzio-Donahue C, Jimeno A, Gabrielson KL, Matsui W, Maitra A
Cancer Res., 2007-03-01;67(5):2187-96.
Species: Human
Sample Types: Whole Cells
Applications: ICC

Expression of hedgehog proteins in the human thymus.
Authors: Sacedon R, Varas A, Hernandez-Lopez C, Gutierrez-deFrias C, Crompton T, Zapata AG, Vicente A
J. Histochem. Cytochem., 2003-11-01;51(11):1557-66.
Species: Human
Sample Types: Whole Tissue
Applications: IHC-Fr