Anti-Smad3 (phospho S423 + S425) Rabbit mAb - ChIP, CUT&RUN and CUT&Tag Grade

Product

Anti-Smad3 (phospho S423 + S425) Rabbit mAb - ChIP, CUT&RUN and CUT&Tag Grade

Catalog number :AT0649

Members of the Smad family of signal transduction molecules are components of a critical intracellular pathway that transmit TGF-β signals from the cell surface into the nucleus. Three distinct classes of Smads have been defined: the receptor-regulated Smads (R-Smads), which include Smad1, 2, 3, 5, and 8; the common-mediator Smad (co-Smad), Smad4; and the antagonistic or inhibitory Smads (I-Smads), Smad6 and 7. Activated type I receptors associate with specific R-Smads and phosphorylate them on a conserved carboxy terminal SSXS motif. The phosphorylated R-Smad dissociates from the receptor and forms a heteromeric complex with the co-Smad (Smad4), allowing translocation of the complex to the nucleus. Once in the nucleus, Smads can target a variety of DNA binding proteins to regulate transcriptional responses.

Following stimulation by TGF-β, Smad2 and Smad3 become phosphorylated at carboxyl terminal serine residues (Ser465 and 467 on Smad2; Ser423 and 425 on Smad3) by TGF-β Receptor I. Phosphorylated Smad 2/3 can complex with Smad4 and translocate to the nucleus to regulate gene expression.

Overview

Reactivity: Mouse, Human

Tested applications: WB : 1/500-1/2000. Predicted molecular weight: 55kDa.

IHC : 1/50~1/200

IF/ICC : 1/50~1/200

FC : 1/50
ChIP, CUT&RUN and CUT&Tag

Not yet tested in other applications. Optimal dilutions/concentrations should be determined by the end user.

Specificity: This antibody detects endogenous levels of Smad3 when phosphorylated at Ser423/425. This antibody does not cross-react with other family members.

Properties

Immunogen: Anti-SMAD3 pS423pS425 antibody was prepared from whole rabbit serum produced by repeated immunizations with a dual phosphorylated synthetic peptide corresponding to a c-terminal region with Serine 423 and Serine 425 of human Smad3 protein.

Clonality: Monoclonal, clone number: 6A2

Isotype: Rabbit IgG

Form: Liquid, 50 μl,1mg/ml, PBS (pH 7.2) and 40% Glycerol,0.02% Sodium Azide

Storage instruction: store at -20°C, Avoid freeze / thaw cycle.

Host: Rabbit

Applications

WB Image: Western blot – Smad3 (phospho S423 + S425) antibody (AT0649) at 1/1000 dilution

Sample: in A549 cell lysate treated with TGF-ß1

Observed band size: 55 kDa

Recommended secondary antibody: Goat anti rabbit IgG-HRP (Engibody, Catalog number:AT0097)

Recommended ECL: ECL Pico-Detect™ Western Chemiluminescent HRP Substrate (Engibody, Catalog number: IF6747)

IHC Image: Immunohistochemistry: Smad3 [p Ser423, p Ser425] Antibody - Used at 1:100 to detect signal in a variety of tissues including multi-human, multi-brain and multi-cancer slides. This image shows strong nuclear staining in the majority of epidermal keratinocytes at 40X. Tissue was formalin-fixed and paraffin embedded. The image shows localization of the antibody as the precipitated red signal, with a hematoxylin purple nuclear counterstain.

ICC/IF Image: Immunocytochemistry/Immunofluorescence: Smad3 [p Ser423, p Ser425] Antibody - Staining Smad3[p Ser423, p Ser425] in A549 cells treated with TGFβ (green). The nuclear counter stain is DAPI (blue). Cells were fixed in paraformaldehyde, permeabilised with 0.25% Triton X100/PBS.

Anti-Smad3 (phospho S423 + S425) Rabbit mAb - ChIP, CUT&RUN and CUT&Tag Grade

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