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Rabbit Recombinant Monoclonal SMAD4 antibody - conjugated to HRP. Suitable for WB and reacts with Human samples.

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Images

Western blot - HRP Anti-Smad4 antibody [EP618Y] (AB195554), expandable thumbnail
  • Western blot - HRP Anti-Smad4 antibody [EP618Y] (AB195554), expandable thumbnail

Key facts

Isotype
IgG
Host species
Rabbit
Conjugation
HRP
Storage buffer

pH: 7.4
Preservative: 0.1% Proclin 300 Solution
Constituents: PBS, 30% Glycerol (glycerin, glycerine), 1% BSA

Form
Liquid
Clonality
Monoclonal

Immunogen

  • The exact immunogen used to generate this antibody is proprietary information.

Reactivity data

Select an application
Product promiseTestedExpectedPredictedNot recommended
IHC-PWB
Human
Not recommended
Tested
Mouse
Not recommended
Predicted
Rat
Not recommended
Predicted

Not recommended
Not recommended

Species
Human
Dilution info
-
Notes

Perform heat-mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

Species
Mouse
Dilution info
-
Notes

-

Species
Rat
Dilution info
-
Notes

-

Tested
Tested

Species
Human
Dilution info
1/5000
Notes

-

Predicted
Predicted

Species
Mouse, Rat
Dilution info
-
Notes

-

Associated Products

Select an associated product type

5 products for Alternative Product

1 product for Alternative Version

Target data

Function

In muscle physiology, plays a central role in the balance between atrophy and hypertrophy. When recruited by MSTN, promotes atrophy response via phosphorylated SMAD2/4. MSTN decrease causes SMAD4 release and subsequent recruitment by the BMP pathway to promote hypertrophy via phosphorylated SMAD1/5/8. Acts synergistically with SMAD1 and YY1 in bone morphogenetic protein (BMP)-mediated cardiac-specific gene expression. Binds to SMAD binding elements (SBEs) (5'-GTCT/AGAC-3') within BMP response element (BMPRE) of cardiac activating regions (By similarity). Common SMAD (co-SMAD) is the coactivator and mediator of signal transduction by TGF-beta (transforming growth factor). Component of the heterotrimeric SMAD2/SMAD3-SMAD4 complex that forms in the nucleus and is required for the TGF-mediated signaling (PubMed:25514493). Promotes binding of the SMAD2/SMAD4/FAST-1 complex to DNA and provides an activation function required for SMAD1 or SMAD2 to stimulate transcription. Component of the multimeric SMAD3/SMAD4/JUN/FOS complex which forms at the AP1 promoter site; required for synergistic transcriptional activity in response to TGF-beta. May act as a tumor suppressor. Positively regulates PDPK1 kinase activity by stimulating its dissociation from the 14-3-3 protein YWHAQ which acts as a negative regulator.

Alternative names

Recommended products

Rabbit Recombinant Monoclonal SMAD4 antibody - conjugated to HRP. Suitable for WB and reacts with Human samples.

Key facts

Isotype
IgG
Conjugation
HRP
Form
Liquid
Clonality
Monoclonal
Immunogen
  • The exact immunogen used to generate this antibody is proprietary information.
Clone number
EP618Y
Purification technique
Affinity purification Protein A
Concentration
Loading...

Storage

Shipped at conditions
Blue Ice
Appropriate short-term storage duration
1-2 weeks
Appropriate short-term storage conditions
+4°C
Appropriate long-term storage conditions
-20°C
Aliquoting information
Upon delivery aliquot
Storage information
Avoid freeze / thaw cycle, Store in the dark

Notes

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

Supplementary info

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

Smad4 also known as DPC4 or MADH4 is a central protein in the TGF-beta signaling pathway with a molecular mass of approximately 60 kDa. It acts as a signal transducer that facilitates communication from the cell surface to the nucleus. Smad4 is broadly expressed in various tissues playing an important role in the regulation of cellular processes. It forms a complex with receptor-regulated Smads (R-Smads) to translocate to the nucleus where it influences gene transcription.

Biological function summary

Smad4 influences cell proliferation differentiation and apoptosis by mediating signals from TGF-beta cytokines. It is part of the Smad protein family acting as a transcriptional controller. Upon TGF-beta receptor activation Smad4 forms complexes with Smad2 and Smad3 translocating to the nucleus to regulate genes imperative for cellular homeostasis. Its role in cell cycle regulation underlines its contribution to normal cellular functions and its potential involvement in disorders.

Pathways

Smad4 operates within the TGF-beta pathway linking extracellular signals to nuclear transcription alterations. It participates in the regulation of epithelial-mesenchymal transition (EMT) a process important for development and tumor progression. In these pathways Smad4 interacts closely with Smad2 and Smad3 orchestrating various cellular responses to external stimuli through transcriptional management.

Associated diseases and disorders

Smad4 is highly related to cancer and juvenile polyposis syndrome. Mutations or deletions in Smad4 disrupt its function contributing to the progression of pancreatic cancer and colorectal cancer among others. Within these contexts Smad4 connects strongly to other proteins like p21 and cyclin-dependent kinase inhibitors which are important in cell cycle arrest and impede tumor growth.

Product promise

We are dedicated to supporting your work with high quality reagents and we are here for you every step of the way should you need us.

In the unlikely event of one of our products not working as expected, you are covered by our product promise.

Full details and terms and conditions can be found here:
Terms & Conditions.

2 product images

  • Western blot - HRP Anti-Smad4 antibody [EP618Y] (ab195554), expandable thumbnail

    Western blot - HRP Anti-Smad4 antibody [EP618Y] (ab195554)

    ab195554 was shown to recognize Smad4 in wild-type HAP1 cells as signal was lost at the expected MW in SMAD4 knockout cells. Additional cross-reactive bands were observed in the wild-type and knockout cells. Wild-type and SMAD4 knockout samples were subjected to SDS-PAGE. ab195554 and Alexa Fluor® 680 Anti-GAPDH antibody [mAbcam 9484] - Loading Control ab184095 (Mouse monoclonal [mAbcam 9484] to GAPDH - Loading Control (Alexa Fluor® 680) loading control) were incubated overnight at 4°C at 1/5000 dilution and 1/20000 dilution respectively. The loading control was imaged using the Licor Odyssey CLx prior to blots being developed with ECL technique.

    All lanes: Western blot - HRP Anti-Smad4 antibody [EP618Y] (ab195554) at 1/5000 dilution

    Lane 1: Wild-type HAP1 whole cell lysate at 20 µg

    Lane 2: SMAD4 knockout HAP1 whole cell lysate at 20 µg

    Predicted band size: 60 kDa

    Observed band size: 65 kDa

    Exposure time: 12min

  • Western blot - HRP Anti-Smad4 antibody [EP618Y] (ab195554), expandable thumbnail

    Western blot - HRP Anti-Smad4 antibody [EP618Y] (ab195554)

    This blot was produced using a 4-12% Bis-tris gel under the MOPS buffer system. The gel was run at 200V for 50 minutes before being transferred onto a Nitrocellulose membrane at 30V for 70 minutes. The membrane was then blocked for an hour using 3% milk before being incubated with ab195554 overnight at 4°C. Antibody binding was visualised using ECL development solution ECL Substrate Kit (High Sensitivity) ab133406.

    All lanes: Western blot - HRP Anti-Smad4 antibody [EP618Y] (ab195554) at 1/5000 dilution

    All lanes: SHSY-5Y (Human neuroblastoma cell line) Whole Cell Lysate at 10 µg

    Developed using the ECL technique.

    Performed under reducing conditions.

    Predicted band size: 60 kDa, 82 kDa

    Observed band size: 30 kDa, 47 kDa, 52 kDa, 54 kDa, 65 kDa, 82 kDa

    Exposure time: 20min

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Product protocols

For this product, it's our understanding that no specific protocols are required. You can:

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