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Anti-SHH Antibody EP1190Y
|A synthetic peptide corresponding to residues within c-product subunit of human SHH protein was used as an immunogen. This antibody is specific for both full length (50 kDa) and c-product subunit (27 kDa) of human SHH protein.|
||0.5~1.0 mg/ml (Lot Dependent)
|WB, IHC, FC
||WB: 1:1000 - 1:10000; IP: 1:10 - 1:100; FC: 1:10 - 1:100; IHC-P: 1:100
|PBS 49%,Sodium azide 0.01%,Glycerol 50%,BSA 0.05%|
|Tissue culture supernatant (Protein A or G Sepharose)
|Does not react with Mouse, Rat
|Homo sapiens sonic hedgehog (SHH)|
|HHG1; HLP3; HPE3; MCOPCB5; SMMCI; TPT; TPTPS|
|Sonic Hedgehog Protein (SHH, VHH-1) belongs to hedgehog protein family which includes Indian Hh, and Desert Hh. Hh family is involved in the cell fate and patterning during embryonic development, homeostasis, and adult tissue renewal (1). Similar to other member in the family, SHH binds to the patched (PTC) cell surface receptor, releasing the signal transducer Smoothened (Smo) to transmit the Hh signal into the cell and activate transcription of the target gene (2). Precursor SHH is autocatlytically cleaved into two subunits, N-terminal and C-terminal products. Soluble N-terminal product is involved in the signaling activity, while C-product displays an autoproteolysis activity on the precursor and a cholesterol transferase activity on the N-terminal product. C-terminal product attaches a cholesterol moiety to the N-terminal product, preventing N-terminal product diffusion within the developing embryo (3). A defect in SHH has been linked in holoprosencephaly type 3 (HPE3), in which developing forebrain fails to separate into right and left hemispheres, and ocular coloboma (4). |
Hedgehog signaling pathway
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Western blot - Sonic Hedgehog antibody [EP1190Y]; Anti-Sonic Hedgehog antibody [EP1190Y] 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.
Immunohistochemistry (Paraffin-embedded sections) - Sonic Hedgehog antibody [EP1190Y]; Human kidney cancerous tissue labelled with ab53281 at 1/100 dilution
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Sonic Hedgehog antibody [EP1190Y]; 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.
Flow Cytometry-Anti-Sonic Hedgehog antibody [EP1190Y](ab53281); 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 for 30 min at 22°C. The secondary antibody used was DyLight? 488 goat anti-rabbit IgG (H+L) at 1/500 dilution for 30 min at 22°C. Isotype control antibody (black line) was rabbit IgG (monoclonal) (1µg/1x10^6 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.