Sonic Hedgehog/Shh Antibody - C-terminus

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.

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.

• Reactivity: Mu
• Applications: WB, IHC
• Clonality: Polyclonal
• Host: Goat
• Conjugate: Unconjugated
• Concentration: LYOPH

Sonic Hedgehog/Shh Antibody - C-terminus Summary

• Immunogen: E. coli-derived recombinant mouse Sonic Hedgehog
  Cys25-Ser437 with a Lys122Arg substitution
  Accession # Q62226
• 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: N/A
• Isotype: IgG
• Clonality: Polyclonal
• Host: Goat
• Gene: SHH
• Purity: Immunogen affinity purified
• Purity Statement: Antigen Affinity-purified
• Endotoxin Note: <0.10 EU per 1 μg of the antibody by the LAL method.

Applications/Dilutions

• Dilutions:
  • Immunohistochemistry 5-15 ug/mL
  • Western Blot 1 ug/mL

Packaging, Storage & Formulations

• 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.
• Buffer: 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.
• Preservative: No Preservative
• Concentration: LYOPH
• Purity: Immunogen affinity purified
• Reconstitution Instructions: Reconstitute at 0.2 mg/mL in sterile PBS.

Notes

This product is produced by and ships from R&D Systems, Inc., a Bio-Techne brand.

Alternate Names for Sonic Hedgehog/Shh Antibody - C-terminus

• 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

Background

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.

Limitations

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Primary Antibodies are guaranteed for 1 year from date of receipt.

Publications for Sonic Hedgehog/Shh Antibody (AF445)(8)

Publications using AF445

• A Mafi, R Purohit, E Vielmas, AR Lauinger, B Lam, YS Cheng, T Zhang, Y Huang, SK Kim, WA Goddard, AE Ondrus Hedgehog proteins create a dynamic cholesterol interface PLoS ONE, 2021-02-25;16(2):e0246814. 2021-02-25 [PMID: 33630857] (Western Blot, Human)
• A Andreu-Cer, I Anselme, A Karam, C Laclef, M Catala, S Schneider- The ciliopathy gene ftm/rpgrip1l controls mouse forebrain patterning via region-specific modulation of hedgehog/gli signaling J. Neurosci., 2019-01-28;0(0):. 2019-01-28 [PMID: 30692221] (IHC, Mouse)
• 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 Loss of the transcription factor GLI1 identifies a signaling network in the tumor microenvironment mediating KRAS oncogene-induced transformation. J Biol Chem, 2013-03-12;288(17):11786-94. 2013-03-12 [PMID: 23482563] (IHC-P, Mouse)
• Irvin DK, Jouanneau E, Duvall G T cells enhance stem-like properties and conditional malignancy in gliomas. PLoS ONE, 2010-06-07;5(6):e10974. 2010-06-07 [PMID: 20539758] (Flow Cytometry, Mouse)
• Nolan-Stevaux O, Lau J, Truitt ML, Chu GC, Hebrok M, Fernandez-Zapico ME, Hanahan D GLI1 is regulated through Smoothened-independent mechanisms in neoplastic pancreatic ducts and mediates PDAC cell survival and transformation. Genes Dev., 2009-01-01;23(1):24-36. 2009-01-01 [PMID: 19136624] (IHC-P, Mouse)
• Vue TY, Aaker J, Taniguchi A Characterization of progenitor domains in the developing mouse thalamus. J. Comp. Neurol., 2007-11-01;505(1):73-91. 2007-11-01 [PMID: 17729296] (IHC-Fr, Mouse)
• 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 Blockade of hedgehog signaling inhibits pancreatic cancer invasion and metastases: a new paradigm for combination therapy in solid cancers. Cancer Res., 2007-03-01;67(5):2187-96. 2007-03-01 [PMID: 17332349] (ICC, Human)
• Sacedon R, Varas A, Hernandez-Lopez C, Gutierrez-deFrias C, Crompton T, Zapata AG, Vicente A Expression of hedgehog proteins in the human thymus. J. Histochem. Cytochem., 2003-11-01;51(11):1557-66. 2003-11-01 [PMID: 14566027] (IHC-Fr, Human)

Bioinformatics

• Gene Symbol: SHH
• Uniprot:
  • Q62226()