Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EP1190Y] to Sonic Hedgehog
- Suitable for: ICC/IF, WB, Flow Cyt, IHC-P
- Reacts with: Human
Product nameAnti-Sonic Hedgehog antibody [EP1190Y]
See all Sonic Hedgehog primary antibodies
DescriptionRabbit monoclonal [EP1190Y] to Sonic Hedgehog
SpecificitySpecific for both full length (51kDa) and c-product subunit (27kDa) of human Sonic Hedgehog protein
Tested applicationsSuitable for: ICC/IF, WB, Flow Cyt, IHC-Pmore details
Species reactivityReacts with: Human
Synthetic peptide within Human Sonic Hedgehog. The exact sequence is proprietary.
(Peptide available as
- Human fetal liver and kidney lysates, human kidney, fetal membrane and human pancreatic carcinoma tissue and HeLa cells.
This product is a recombinant monoclonal antibody, which offers several advantages including:
- - High batch-to-batch consistency and reproducibility
- - Improved sensitivity and specificity
- - Long-term security of supply
- - Animal-free production
Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.
We are constantly working hard to ensure we provide our customers with best in class antibodies. As a result of this work we are pleased to now offer this antibody in purified format. We are in the process of updating our datasheets. The purified format is designated 'PUR' on our product labels. If you have any questions regarding this update, please contact our Scientific Support team.
Alexa Fluor® is a registered trademark of Molecular Probes, Inc, a Thermo Fisher Scientific Company. The Alexa Fluor® dye included in this product is provided under an intellectual property license from Life Technologies Corporation. As this product contains the Alexa Fluor® dye, the purchase of this product conveys to the buyer the non-transferable right to use the purchased product and components of the product only in research conducted by the buyer (whether the buyer is an academic or for-profit entity). As this product contains the Alexa Fluor® dye the sale of this product is expressly conditioned on the buyer not using the product or its components, or any materials made using the product or its components, in any activity to generate revenue, which may include, but is not limited to use of the product or its components: in manufacturing; (ii) to provide a service, information, or data in return for payment (iii) for therapeutic, diagnostic or prophylactic purposes; or (iv) for resale, regardless of whether they are sold for use in research. For information on purchasing a license to this product for purposes other than research, contact Life Technologies Corporation, 5781 Van Allen Way, Carlsbad, CA 92008 USA or firstname.lastname@example.org.
Storage instructionsShipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C. Avoid freeze / thaw cycle. Stable for 12 months at -20°C.
Storage bufferpH: 7.20
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol, 0.05% BSA
Concentration information loading...
PurityProtein A purified
Immunizing Peptide (Blocking)
Our Abpromise guarantee covers the use of ab53281 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|ICC/IF||1/250 - 1/500.|
|WB||1/1000 - 1/10000. Detects a band of approximately 51 kDa (predicted molecular weight: 51 kDa).Can be blocked with Sonic Hedgehog peptide (ab203144).|
|Flow Cyt||1/10 - 1/100.
ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.
|IHC-P||1/2000. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
For unpurified use at 1/100.
FunctionBinds to the patched (PTC) receptor, which functions in association with smoothened (SMO), to activate the transcription of target genes. In the absence of SHH, PTC represses the constitutive signaling activity of SMO. Also regulates another target, the gli oncogene. Intercellular signal essential for a variety of patterning events during development: signal produced by the notochord that induces ventral cell fate in the neural tube and somites, and the polarizing signal for patterning of the anterior-posterior axis of the developing limb bud. Displays both floor plate- and motor neuron-inducing activity. The threshold concentration of N-product required for motor neuron induction is 5-fold lower than that required for floor plate induction.
Tissue specificityExpressed in fetal intestine, liver, lung, and kidney. Not expressed in adult tissues.
Involvement in diseaseDefects in SHH are the cause of microphthalmia isolated with coloboma type 5 (MCOPCB5) [MIM:611638]. Microphthalmia is a clinically heterogeneous disorder of eye formation, ranging from small size of a single eye to complete bilateral absence of ocular tissues. Ocular abnormalities like opacities of the cornea and lens, scaring of the retina and choroid, cataract and other abnormalities like cataract may also be present. Ocular colobomas are a set of malformations resulting from abnormal morphogenesis of the optic cup and stalk, and the fusion of the fetal fissure (optic fissure).
Defects in SHH are the cause of holoprosencephaly type 3 (HPE3) [MIM:142945]. Holoprosencephaly (HPE) [MIM:236100] is the most common structural anomaly of the brain, in which the developing forebrain fails to correctly separate into right and left hemispheres. Holoprosencephaly is genetically heterogeneous and associated with several distinct facies and phenotypic variability. The majority of HPE3 cases are apparently sporadic, although clear examples of autosomal dominant inheritance have been described. Interestingly, up to 30% of obligate carriers of HPE3 gene in autosomal dominant pedigrees are clinically unaffected.
Defects in SHH are a cause of solitary median maxillary central incisor (SMMCI) [MIM:147250]. SMMCI is a rare dental anomaly characterized by the congenital absence of one maxillary central incisor.
Defects in SHH are the cause of triphalangeal thumb-polysyndactyly syndrome (TPTPS) [MIM:174500]. TPTPS is an autosomal dominant syndrome characterized by a wide spectrum of pre- and post-axial abnormalities due to altered SHH expression pattern during limb development. TPTPS mutations have been mapped to the 7q36 locus in the LMBR1 gene which contains in its intron 5 a long-range cis-regulatory element of SHH expression.
Sequence similaritiesBelongs to the hedgehog family.
modificationsThe C-terminal domain displays an autoproteolysis activity and a cholesterol transferase activity. Both activities result in the cleavage of the full-length protein and covalent attachment of a cholesterol moiety to the C-terminal of the newly generated N-terminal fragment (N-product). The N-product is the active species in both local and long-range signaling, whereas the C-product has no signaling activity.
Cholesterylation is required for N-product targeting to lipid rafts and multimerization.
N-palmitoylation of Cys-24 by HHAT is required for N-product multimerization and full activity.
Cellular localizationCell membrane. The N-product either remains associated with lipid rafts at the cell surface, or forms freely diffusible active multimers with its hydrophobic lipid-modified N- and C-termini buried inside and Secreted > extracellular space. The C-terminal peptide diffuses from the cell.
- Information by UniProt
- HHG 1 antibody
- HHG-1 antibody
- HHG1 antibody
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human pancreatic carcinoma tissue sections labeling Sonic Hedgehog with purified ab53281 at 1/2000 dilution (0.097 μg/ml). Heat mediated antigen retrieval was performed using EDTA Buffer, PH9. Tissue was counterstained with Hematoxylin. ab97051 Goat Anti-Rabbit IgG H&L (HRP) secondary antibody was used at 1/500 dilution. PBS instead of the primary antibody was used as the negative control.
Immunocytochemistry/ Immunofluorescence analysis of HeLa (Human epithelial cell line from cervix adenocarcinoma) cells labeling Sonic Hedgehog with purified ab53281 at 1/250 dilution (0.8μg/ml). Cells were fixed in 4% Paraformaldehyde and permeabilized with 0.1% tritonX-100. Cells were counterstained with Ab195889, Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) 1/200 (2.5 μg/ml). ab150077 Goat anti rabbit IgG(Alexa Fluor® 488) was used as the secondary antibody at 1/1000 dilution. DAPI nuclear counterstain. PBS instead of the primary antibody was used as the secondary antibody only control.
Anti-Sonic Hedgehog antibody [EP1190Y] (ab53281) at 1/2000 dilution (purified) + Human fetal liver lysate at 15 µg
Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution
Predicted band size: 51 kDa
Observed band size: 27,50 kDa why is the actual band size different from the predicted?
Blocking and diluting buffer: 5% NFDM/TBST.
50kDa: Full-length SHH; 27KDa: C-terminal product
Flow Cytometry analysis of HeLa (Human epithelial cell line from cervix adenocarcinoma) cells labeling Sonic Hedgehog with purified ab53281 at 1/20 dilution (10 ug/ml) (red). Cells were fixed with 4% Paraformaldehyde. A Goat anti rabbit IgG (Alexa Fluor® 488) secondary antibody was used at 1/2000 dilution. Isotype control - Rabbit monoclonal IgG (Black). Unlabeled control - cells without incubation with primary antibody and secondary antibody (Blue).
Immunocytochemistry/Immunofluorescence analysis of HeLa (human cervix adenocarcinoma) labelling Sonic Hedgehog with purified ab53281 at 1/500. Cells were fixed with 4% Paraformaldehyde and permeabilised by 0.1% tritonX-100. An Alexa Fluor® 488-conjugated goat anti-rabbit IgG (1/1000) was used as the secondary antibody (Ab150077). Nuclei counterstained with DAPI (blue).
Immunocytochemistry/Immunofluorescence analysis of HepG2 cells labelling Sonic Hedgehog with unpurified ab53281.
Anti-Sonic Hedgehog antibody [EP1190Y] (ab53281) at 1/1000 dilution (purified) + Human fetal kidney lysate at 15 µg
Anti-Rabbit IgG (HRP), specific to the non-reduced form of IgG at 1/2000 dilution
Predicted band size: 51 kDa
Observed band size: 27 kDa why is the actual band size different from the predicted?
Blocking and diluting buffer: 5% NFDM /TBST .
27KDa: C-terminal product.
Flow Cytometry analysis of HeLa (human cervix adenocarcinoma) cells labelling Sonic Hedgehog with unpurified ab53281 at 1/20 (red). Cells were fixed with 4% paraformaldehyde and permeabilised with 90% methanol. An Alexa Fluor® 488-conjugated goat anti-rabbit IgG (1/2000) was used as the secondary antibody. Black - Isotype control, rabbit monoclonal IgG. Blue - Unlabelled control, cells without incubation with primary and secondary antibodies.
Anti-Sonic Hedgehog antibody [EP1190Y] (ab53281) at 1/2000 dilution (unpurified) + Fetal liver membrane at 10 µg
Goat anti-rabbit HRP at 1/2000 dilution
Predicted band size: 51 kDa
Observed band size: 51 kDa
Additional bands at: 27 kDa (possible isoform), 49 kDa. We are unsure as to the identity of these extra bands.
Immunohistochemical analysis of Formalin fixed paraffin-embedded human kidney cancerous tissue labeling Sonic Hedgehog with unpurified ab53281 at 1/100 dilution.
Immunohistochemical analysis of Formalin fixed paraffin embedded human ovarian cancer tissue, staining Sonic Hedgehog with unpurified ab53281.
Antigen retrieval was carried out in citrate buffer using a pressure cooker for 40 minutes. Sections were blocked with blocking agent before incubating with primary antibody (1/2000) for 90 minutes at room temperature. Staining was detected using DAB.
Overlay histogram showing HepG2 cells stained with unpurified ab53281 (red line). The cells were fixed with 4% paraformaldehyde (10 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab53281, 1/100 dilution) for 30 min at 22ºC. The secondary antibody used was DyLight® 488 goat anti-rabbit IgG (H+L) (ab96899) at 1/500 dilution for 30 min at 22ºC. Isotype control antibody (black line) was rabbit IgG (monoclonal) (1µg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed. This antibody gave a positive signal in HepG2 cells fixed with 80% methanol (5 min)/permeabilized with 0.1% PBS-Tween for 20 min used under the same conditions.
To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.
ab53281 has been referenced in 50 publications.
- Qin Y et al. Sonic hedgehog signaling pathway in Myelodysplastic Syndrome: Abnormal activation and jervine intervention. Gene 754:144881 (2020). PubMed: 32526259
- Ke B et al. Sonic Hedgehog/Gli1 Signaling Pathway Regulates Cell Migration and Invasion via Induction of Epithelial-to-mesenchymal Transition in Gastric Cancer. J Cancer 11:3932-3943 (2020). PubMed: 32328197
- Zhou H et al. Chidamide Inhibits Glioma Cells by Increasing Oxidative Stress via the miRNA-338-5p Regulation of Hedgehog Signaling. Oxid Med Cell Longev 2020:7126976 (2020). PubMed: 32256960
- Estep M et al. Hepatic sonic hedgehog protein expression measured by computer assisted morphometry significantly correlates with features of non-alcoholic steatohepatitis. BMC Gastroenterol 19:27 (2019). PubMed: 30744560
- Adams CR et al. Transcriptional control of subtype switching ensures adaptation and growth of pancreatic cancer. Elife 8:N/A (2019). PubMed: 31134896