Rabbit Polyclonal (Mouse)Shh(N-term) Antibody

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     All lanes : Anti-Shh Antibody (N-term) at 1:2000 dilution Lane 1: F9 whole cell lysate Lane 2: mouse stomach lysates Lane 3: rat liver whole cell lysates Lane 4: NIH/3T3 lysates Lysates/proteins at 20 µg per lane. Secondary Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated at 1/10000 dilution Predicted band size : 48 kDa Blocking/Dilution buffer: 5% NFDM/TBST. Lysates/proteins at 20 µg per lane. Secondary Goat Anti-Rabbit IgG,  (H+L), Peroxidase conjugated at 1/10000 dilution. Predicted band size : 84 kDa Blocking/Dilution buffer: 5% NFDM/TBST.    

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     All lanes : Anti-Shh Antibody (N-term) at 1:2000 dilution Lane 1: F9 whole cell lysates Lane 2: mouse stomach lysates Lane 3: NIH/3T3 whole cell lysates Lane 4: rat liver lysates Lysates/proteins at 20 µg per lane. Secondary Goat Anti-Rabbit IgG,  (H+L), Peroxidase conjugated at 1/10000 dilution Predicted band size : 48 kDa Blocking/Dilution buffer: 5% NFDM/TBST.    

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     Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized Hela (Human Cervical epithelial adenocarcinoma cell line) cells labeling Shh with P34284 at 1/25 dilution, followed by Dylight® 488-conjugated goat anti-rabbit IgG  secondary antibody at 1/200 dilution (green). Immunofluorescence image showing cytoplasm and membrane staining on Hela cell line. Cytoplasmic actin is detected with Dylight® 554 Phalloidin  at 1/100 dilution (red).The nuclear counter stain is DAPI (blue).    

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     Overlay histogram showing HT-29 cells stained with P34284 (green line).  The cells were fixed with 4% paraformaldehyde (10 min) and then permeabilized with 90% methanol for 10 min.  The cells were then icubated in 2% bovine serum albumin to block non-specific protein-protein interactions followed by the antibody (AP12735b,  1:25 dilution) for 60 min at 37ºC.  The secondary antibody used was Alexa Fluor® 488 goat anti-rabbit lgG (H+L) (1583138) at 1/400 dilution for 40 min at 37ºC.  Isotype control antibody (blue line) was rabbit IgG1 (1μg/1x10^6 cells) used under the same conditions.  Acquisition of >10, 000 events was performed.    

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     P34284 staining (Mouse) Shh in mouse cerebellum sections by Immunohistochemistry (IHC-P - paraformaldehyde-fixed,  paraffin-embedded sections).  Tissue was fixed with formaldehyde and blocked with 3% BSA for 0. 5 hour at room temperature; antigen retrieval was by heat mediation with a citrate buffer (pH6).  Samples were incubated with primary antibody (1/25) for 1 hours at 37°C.  A undiluted biotinylated goat polyvalent antibody was used as the secondary antibody.    

The Mouse Shh (N-term) antibody is designed to detect the N-terminal region of Sonic Hedgehog (Shh), a critical morphogen involved in embryonic development, tissue patterning, and cellular differentiation. Shh, a secreted glycoprotein, plays a pivotal role in regulating organogenesis, particularly in the nervous system, limbs, and gut. Its active N-terminal domain mediates signaling by binding to the Patched (Ptch) receptor, relieving suppression of the Smoothened (Smo) receptor and activating downstream transcriptional targets. Dysregulation of Shh signaling is linked to developmental anomalies and cancers, such as medulloblastoma and basal cell carcinoma.

This antibody specifically targets epitopes within the N-terminal region of Shh, enabling researchers to study its expression, processing, and localization in various biological contexts. It is commonly utilized in techniques like Western blotting, immunohistochemistry, and immunofluorescence to investigate Shh dynamics in developmental models, disease mechanisms, or stem cell research. As the N-terminal fragment is essential for Shh's biological activity, the antibody serves as a valuable tool for elucidating its functional roles and interactions within signaling pathways. Validation typically includes reactivity checks across species (e.g., human, mouse) and applications to ensure specificity, supporting its reliability in both basic and translational studies.