RabMAb

Anti-Sonic Hedgehog抗体[EP1190Y] (ab53281)

概述

  • 产品名称Anti-Sonic Hedgehog抗体[EP1190Y]
    参阅全部 Sonic Hedgehog 一抗
  • 描述
    兔单克隆抗体[EP1190Y] to Sonic Hedgehog
  • 特异性Specific for both full length (51kDa) and c-product subunit (27kDa) of human Sonic Hedgehog protein
  • 经测试应用适用于: ICC/IF, WB, Flow Cyt, IHC-Pmore details
  • 种属反应性
    与反应: Human
  • 免疫原

    Synthetic peptide (Human) corresponding to residues within c-product subunit

  • 阳性对照
    • Fetal liver membrane Human kidney tissue FC: HeLa cells
  • 常规说明

    Produced using Abcam's RabMAb® technology. RabMAb® technology is covered by the following U.S. Patents, No. 5, 675, 063 and/or 7, 429, 487.

    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 use products containing Alexa Fluor® dyes for purposes other than research, contact Life Technologies Corporation, 5791 Van Allen Way, Carlsbad, CA 92008 USA or outlicensing@lifetech.com

应用

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.

应用 Ab评论 说明
ICC/IF 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 1/100.

靶标

  • 功能Binds 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.
  • 组织特异性Expressed in fetal intestine, liver, lung, and kidney. Not expressed in adult tissues.
  • 疾病相关Defects 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.
  • 序列相似性Belongs to the hedgehog family.
  • 翻译后修饰The 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.
  • 细胞定位Cell 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
  • 数据库链接
  • 别名
    • HHG 1 antibody
    • HHG-1 antibody
    • HHG1 antibody
    • HLP 3 antibody
    • HLP3 antibody
    • Holoprosencephaly 3 antibody
    • HPE 3 antibody
    • HPE3 antibody
    • MCOPCB5 antibody
    • shh antibody
    • SHH_HUMAN antibody
    • SMMC I antibody
    • SMMCI antibody
    • Sonic Hedgehog (Drosophila) homolog antibody
    • sonic hedgehog homolog (Drosophila) antibody
    • Sonic hedgehog homolog antibody
    • Sonic hedgehog protein antibody
    • Sonic hedgehog protein C-product antibody
    • TPT antibody
    • TPTPS antibody
    see all

Anti-Sonic Hedgehog antibody [EP1190Y] 图像

  • 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).

  • Immunofluorescent staining of HepG2 using anti-Sonic Hedgehog Protein Alexa Fluor 555 RabMAB

  • Flow Cytometry analysis of HeLa (human cervix adenocarcinoma) cells labelling Sonic Hedgehog with 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 + Fetal liver membrane at 10 µg

    Secondary
    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.
  • Human kidney cancerous tissue labelled with ab53281 at 1/100 dilution
  • Immunohistochemical analysis of Human ovarian cancer tissue, staining Sonic Hedgehog with 53281.

    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 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.

Anti-Sonic Hedgehog antibody [EP1190Y] (ab53281)参考文献

This product has been referenced in:
  • Verrill C  et al. Altered expression of epithelial-to-mesenchymal transition proteins in extraprostatic prostate cancer. Oncotarget 7:1107-19 (2016). Read more (PubMed: 26701730) »
  • Zhang X  et al. Expression of SOX9 and CDX2 in nongoblet columnar-lined esophagus predicts the detection of Barrett's esophagus during follow-up. Mod Pathol 28:654-61 (2015). Read more (PubMed: 25412842) »

See all 15 Publications for this product

Product Wall

Application Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample Human Tissue sections (tooth)
Antigen retrieval step None
Permeabilization No
Specification tooth
Blocking step BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
Fixative Paraformaldehyde
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提交于 Nov 19 2015

Application Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample Human Tissue sections (pancreatic tumor)
Antigen retrieval step Heat mediated - Buffer/Enzyme Used: 10mM Citrate buffer pH 6
Permeabilization No
Specification pancreatic tumor
Blocking step Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 21°C
Fixative 10% buffered formalin
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提交于 Sep 14 2015

Application Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample Mouse Tissue sections (Pancreatic nodule)
Antigen retrieval step Heat mediated - Buffer/Enzyme Used: 10mM Citrate buffer pH 6
Permeabilization No
Specification Pancreatic nodule
Blocking step Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 21°C
Fixative 10% buffered Formalin
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提交于 Aug 07 2015

Application Western blot
Loading amount 50 µg
Gel Running Conditions Reduced Denaturing
Sample Human Cell lysate - whole cell (fibroblasts)
Specification fibroblasts
Blocking step BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: RT°C
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提交于 Mar 27 2015

The antibody is working in the positive controls but having low expression in your samples. There could be number of reason why lower signal is found in sample tissues. It may be that the expression levels of protein in this sample type are lower than ...

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Abcam guarantees this product to work in the species/application used in this Abreview.
Application Western blot
Sample Human Cell lysate - whole cell (Glioblastoma primary cells)
Loading amount 30 µg
Specification Glioblastoma primary cells
Gel Running Conditions Reduced Denaturing (10%)
Blocking step Milk as blocking agent for 30 minute(s) · Concentration: 10% · Temperature: 20°C
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提交于 Nov 21 2012

Thank you for contacting us. I've listed the recommended running conditions below.

For ab7970: IHC-P: 1/1000 - 1/4000. Perform heat mediated antigen retrieval via the microwave method before commencing with IHC staining protocol.

F...

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Thank you for confirming these details and for your cooperation. The details provided enable us to closely monitor the quality of our products.

I am sorry this product did not perform as stated on the datasheet and for the inconvenience this...

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Thank you for bringing this to our attention. This is the first complaint we have received for this antibody and it sells very well. I suspect the single vial may have become contaminated. Have you stored it as specified on the datasheet, at -20C?
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Thank you for contacting Abcam regarding ab53281.


The immunogen used to generate this antibody is from the C-product, however the antibody will recognize precursor SHH as well which will account for the membrane staining in FACS. If you w...

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