Anti-Smoothened Rabbit mAb

Product

Anti-Smoothened Rabbit mAb

Catalog number :AT0402

G protein-coupled receptor that probably associates with the patched protein (PTCH) to transduce the hedgehog's proteins signal. Binding of sonic hedgehog (SHH) to its receptor patched is thought to prevent normal inhibition by patched of smoothened (SMO). Required for the accumulation of KIF7 and GLI3 in the cilia.

Overview

Description: Rabbit monoclonal antibody to Smoothened

Reactivity: Mouse, Human, Monkey, Zebrafish

Tested applications: WB: Use a concentration of 0.1 - 1 µg/ml. Detects a band of approximately 86 kDa (predicted molecular weight: 86 kDa).

IHC-P: Use a concentration of 10 µg/ml.

ICC/IF: Use at an assay dependent concentration.

Properties

Immunogen: Synthetic peptide (Human) corresponding to a region between residues 190-240 of human Smoothened. (NP_005622.1).

Clonality: Monoclonal, clone number: 23B7

Isotype: Rabbit IgG

Form: Liquid
Preservative: 0.05% Sodium Azide
Constituents: 0.05% BSA, PBS

Storage instruction: Shipped at 4°C. Upon delivery aliquot and store at -20°C or -80°C. Avoid repeated freeze / thaw cycles.

Database links

Applications

WB Image: All lanes : Anti-Smoothened antibody (AT0402) at 0.1 µg/ml

Lane 1 : Human heart lysate
Lane 2 : Human heart lysate with immunising peptide

Predicted band size : 86 kDa
Observed band size : 86 kDa

IHC Image: Immunohistochemical analysis using formalin/PFA-fixed paraffin-embedded human brain tissue with an isotype control (top image) and AT0402, at 10µg/ml staining Smoothened (bottom image).
AT0402 staining Smoothened in 6 month-old transgenic zebrafish pancreas (Ihha overexpression) by Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections).

Sections were incubated with primary antibody (1/200) and HRP-conjugated secondary antibody colored using DAB solution. Slides were counterstained with hematoxylin.

ICC/IF Image: ICC/IF image of AT0402 stained HeLa cells. The cells were 4% formaldehyde fixed (10 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (AT0402, 1µg/ml) overnight at +4°C. The secondary antibody (green) was Alexa Fluor® 488 goat anti-rabbit IgG (H+L) used at a 1/1000 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM.

Anti-Smoothened Rabbit mAb

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