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AB321949

BOND RX - Anti-Smad3 (phospho S423 + S425) antibody [EP823Y]

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Rabbit Recombinant Monoclonal SMAD3 (pS423/425) + SMAD5 (pS463/465 ) + SMAD1 (pS463/465) + SMAD2 (pS465/467) antibody. Suitable for detection of SMAD3 + SMAD5 + SMAD2 in IHC. Suitable for human and mouse samples. This product is a bundle of 2 x BOND Titration Containers (with 6mL inserts) along with 100 uL vial of Anti-Smad3 (phospho S423 + S425) antibody [EP823Y] - ab52903.

- Validated on the Leica BOND RX automated IHC staining platform for SMAD3 + SMAD5 + SMAD2 IHC
- Specificity and sensitivity confirmed in IHC with multi-tissue microarray (TMA) validation
- EP823Y clone cited in over 570 publications
- Using biophysical QC, antibody identity is confirmed at a molecular level for unrivaled batch-batch consistency

View Alternative Names

MADH3, SMAD3, Mothers against decapentaplegic homolog 3, MAD homolog 3, Mad3, Mothers against DPP homolog 3, hMAD-3, JV15-2, SMAD family member 3, SMAD 3, Smad3, hSMAD3

1 Images
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - BOND RX - Anti-Smad3 (phospho S423 + S425) antibody [EP823Y] (AB321949)
  • IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - BOND RX - Anti-Smad3 (phospho S423 + S425) antibody [EP823Y] (AB321949)

Immunohistochemical analysis of paraffin-embedded human colon tissue labelling SMAD3 phospho S423 + S425 with ab52903 [EP823Y] at 1.5 µg/mL concentration.

All steps were performed at room temperature unless otherwise stated.

Staining was performed on the BOND RX Advanced Staining Instrument from Leica Biosystems, using the BOND Polymer Refine Detection kit (DS9800) and the following protocols :

  1. Deparaffinization - Dewax
  2. Antigen retrieval - HIER with ER2 (pH 9.0), 20 minutes, 100°C
  3. Staining – modified IHC Protocol F (Marker : ab52903 for 15 minutes with 3 x BOND Wash Solution steps) where standard post-primary step has been excluded.

Sections were counterstained with Hematoxylin (DS9800).

Secondary Antibody only control was performed with the same staining protocols, with marker as negative.

Key facts

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Reactivity data

{ "title": "Reactivity Data", "filters": { "stats": ["", "Reactivity", "Dilution Info", "Notes"] }, "values": { "IHC-P": { "reactivity":"TESTED_AND_REACTS", "dilution-info":"1.5 µg/mL", "notes":"<p></p> Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol." } } }
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Product details

This product is a bundle of 2 x BOND Titration Containers (with 6mL inserts) along with 100 uL vial of Anti-Smad3 (phospho S423 + S425) antibody [EP823Y] - ab52903.

Additional Titration Inserts can be purchased from Leica Biosystems, using OPT9719 (box of 50). Or Titration Container/Insert Kit, using OPT9049 (box of 10/50).

Patented technology
Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.

What are the advantages of a recombinant monoclonal antibody?
This product is a recombinant monoclonal antibody, which offers several advantages including:

  • - High batch-to-batch consistency and reproducibility
  • - Improved sensitivity and specificity
  • - Long-term security of supply
  • - Animal-free batch production

For more information, read more on recombinant antibodies.

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What's included?

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Properties and storage information

Shipped at conditions
Blue Ice
Appropriate short-term storage conditions
-20°C
Appropriate long-term storage conditions
-20°C
Storage information
-20°C
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Supplementary information

This supplementary information is collated from multiple sources and compiled automatically.

Smad3 also known as Mothers against decapentaplegic homolog 3 is a protein that plays a mechanical role in signal transduction. It acts mainly as a transcription factor and gets activated through phosphorylation. The molecular weight of Smad3 is approximately 48 kDa. It is expressed widely across numerous tissues including the cellular nucleus where it executes its function after activation.
Biological function summary

Smad3 acts as a mediator of signal transduction for the TGF-beta (transforming growth factor-beta) superfamily forming a complex with phosphorylated Smad2. This enables it to regulate transcriptional activity influencing cell proliferation differentiation and apoptosis. Smad3 also participates in various cellular processes by interacting with other co-factors and regulatory proteins that aid in fine-tuning its function.

Pathways

Smad3 plays an important role in the TGF-beta signaling pathway where it works closely with Smad4 to propagate the signal. Upon phosphorylation it forms a complex with co-Smad (Smad4) and moves into the nucleus to influence gene expression. Smad3 is also involved in pathways related to oncogenesis and tissue fibrosis indicating its significant role in cellular regulation and response mechanisms.

Smad3 is associated with fibrotic diseases and cancers particularly in tissues such as the liver and lungs. Altered Smad3 signaling contributes to the pathological process occurring in fibrotic disorders often interacting with Smad4 in these abnormalities. Dysregulated Smad3 expression or mutations can also lead to oncogenic transformations highlighting its critical involvement in disease states.
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Product protocols

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Target data

SMAD3 is a receptor-regulated SMAD that functions as an intracellular signal transducer and transcriptional modulator, activated by TGF-beta and activin type 1 receptor kinases. It binds to the TRE element in promoters of numerous genes regulated by TGF-beta, and upon forming a complex with SMAD4, activates transcription. Additionally, SMAD3 can form a complex with SMAD4, JUN, and FOS at the AP-1/SMAD site to regulate TGF-beta-mediated transcription. SMAD3 may inhibit wound healing by modulating the growth and migration of primary keratinocytes and altering TGF-beta-mediated monocyte chemotaxis, with this effect being potentially hormone-sensitive. Furthermore, SMAD3 is involved in regulating chondrogenesis and osteogenesis and may inhibit early bone fracture healing. It also positively regulates PDPK1 kinase activity by promoting its dissociation from the 14-3-3 protein YWHAQ, which negatively regulates it. This supplementary information is collated from multiple sources and compiled automatically.
See full target information SMAD3 phospho S423 + S425
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websiteProtocolBooklet
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Product promise

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