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Rabbit Recombinant Monoclonal SMAD4 antibody. Suitable for IHC-P, WB and reacts with Human, Mouse, Rat samples. Cited in 170 publications.


Images

Key facts

Isotype

IgG

Host species

Rabbit

Storage buffer

pH: 7.2 - 7.4
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA

Form

Liquid

Clonality

Monoclonal

Immunogen

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

Select an application
Product promiseTestedExpectedPredictedNot recommended
IHC-PIPFlow CytWBICC/IF
Human
Tested
Not recommended
Not recommended
Tested
Not recommended
Mouse
Expected
Not recommended
Not recommended
Tested
Not recommended
Rat
Expected
Not recommended
Not recommended
Tested
Not recommended

Tested
Tested

Species

Human

Dilution info

1/100

Notes

Perform heat-mediated antigen retrieval before commencing with IHC staining protocol.

Expected
Expected

Species

Mouse, Rat

Dilution info

Use at an assay dependent concentration.

Notes

-

Not recommended
Not recommended

Species

Mouse, Rat, Human

Dilution info

-

Notes

-

Not recommended
Not recommended

Species

Mouse

Dilution info

-

Notes

For unpurified use at 1/20.

ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.

Species

Rat

Dilution info

-

Notes

For unpurified use at 1/20.

ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.

Species

Human

Dilution info

-

Notes

For unpurified use at 1/20.

ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.

Tested
Tested

Species

Mouse

Dilution info

1/5000

Notes

-

Species

Rat

Dilution info

1/5000

Notes

-

Species

Human

Dilution info

1/5000

Notes

-

Not recommended
Not recommended

Species

Mouse

Dilution info

-

Notes

For unpurified use at 1/100 - 1/250.

Species

Rat

Dilution info

-

Notes

For unpurified use at 1/100 - 1/250.

Species

Human

Dilution info

-

Notes

For unpurified use at 1/100 - 1/250.

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

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Rabbit Recombinant Monoclonal SMAD4 antibody. Suitable for IHC-P, WB and reacts with Human, Mouse, Rat samples. Cited in 170 publications.

Alternative names

Key facts

Isotype

IgG

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
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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

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.

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.

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.

11 product images

  • Western blot - Anti-Smad4 antibody [EP618Y] (ab40759), expandable thumbnail

    Western blot - Anti-Smad4 antibody [EP618Y] (ab40759)

    Lane 1: Wild type HAP1 whole cell lysate (20 μg)
    Lane 2: SMAD4 knockout HAP1 whole cell lysate (20 μg)
    Lane 3: HepG2 whole cell lysate (20 μg)
    Lane 4: Jurkat whole cell lysate (20 μg)

    Lanes 1 - 4: Merged signal (red and green). Green - ab40759 observed at 60 kDa. Red - loading control, ab9484, observed at 37 kDa.

    ab40759 was shown to specifically react with SMAD4 in wild type HAP1 cells. No band was observed when SMAD4 knockout HAP1 samples were used. Wild-type and SMAD4 knockout samples were subjected to SDS-PAGE. Ab40759 and ab9484 (Mouse anti GAPDH loading control) were incubated overnight at 4°C at 1000 dilution and 1/20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed ab216773 and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed ab216776 secondary antibodies at 1/20000 dilution for 1 hour at room temperature before imaging.

    All lanes: Western blot - Anti-Smad4 antibody [EP618Y] (AB40759)

    Predicted band size: 60 kDa

  • Western blot - Anti-Smad4 antibody [EP618Y] (ab40759), expandable thumbnail

    Western blot - Anti-Smad4 antibody [EP618Y] (ab40759)

    Blocking and diluting buffer: 5% NFDM/TBST.

    All lanes: Western blot - Anti-Smad4 antibody [EP618Y] (AB40759) at 1/5000 dilution

    Lane 1: SH-SY5Y (Human neuroblastoma cell line from bone marrow) cell lysate at 10 µg

    Lane 2: Ramos (Human Burkitt's lymphoma cell line) cell lysate at 10 µg

    Secondary

    All lanes: Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated at 1/1000 dilution

    Predicted band size: 60 kDa

    Observed band size: 60 kDa

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Smad4 antibody [EP618Y] (ab40759), expandable thumbnail

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Smad4 antibody [EP618Y] (ab40759)

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human lung carcinoma tissue labelling Smad4 with unpurified ab40759 at a 1/100 dilution.

    Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

  • Western blot - Anti-Smad4 antibody [EP618Y] (ab40759), expandable thumbnail

    Western blot - Anti-Smad4 antibody [EP618Y] (ab40759)

    Blocking and dilution buffer: 5% NFDM/TBST.

    All lanes: Western blot - Anti-Smad4 antibody [EP618Y] (AB40759) at 1/5000 dilution

    Lane 1: NIH/3T3 (Mouse embyro fibroblast cell line) cell lysate at 20 µg

    Lane 2: Mouse embryo tissue lysate at 20 µg

    Lane 3: Mouse skin tissue lysate at 20 µg

    Lane 4: Mouse lung tissue lysate at 20 µg

    Lane 5: PC-12 (Rat adrenal gland pheochromocytoma cell line) cell lysate at 20 µg

    Lane 6: C6 cell lysate at 20 µg

    Lane 7: Rat skin tissue lysate at 20 µg

    Lane 8: Rat lung tissue lysate at 20 µg

    Secondary

    All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (AB97051) at 1/20000 dilution

    Predicted band size: 60 kDa

    Observed band size: 60 kDa

    Exposure time: 3min

  • Western blot - Anti-Smad4 antibody [EP618Y] (ab40759), expandable thumbnail

    Western blot - Anti-Smad4 antibody [EP618Y] (ab40759)

    Blocking and dilution buffer: 5% NFDM/TBST.

    All lanes: Western blot - Anti-Smad4 antibody [EP618Y] (AB40759) at 1/5000 dilution

    Lane 1: SW480 (Human colorectal adenocarcinoma cell line) cell lysate at 20 µg

    Lane 2: HepG2 (Human liver hepatocellular carcinoma cell line) cell lysate at 20 µg

    Lane 3: Jurkat (Human T cell leukemia cell line from peripheral blood) cell lysate at 20 µg

    Lane 4: Human skin tissue lysate at 20 µg

    Lane 5: Human lung tissue lysate at 20 µg

    Lane 6: Human artery tissue lysate at 20 µg

    Secondary

    All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (AB97051) at 1/20000 dilution

    Predicted band size: 60 kDa

    Observed band size: 60 kDa

    Exposure time: 30s

  • Western blot - Anti-Smad4 antibody [EP618Y] (ab40759), expandable thumbnail

    Western blot - Anti-Smad4 antibody [EP618Y] (ab40759)

    Different batches of ab40759 were tested on Ramos (Human Burkitt's lymphoma B lymphocyte) lysate at 0.7 µg/ml. 15 µg of lysate was loaded in each lane. Bands observed at 60 kDa.

    All lanes: Western blot - Anti-Smad4 antibody [EP618Y] (AB40759)

    Predicted band size: 60 kDa

  • Western blot - Anti-Smad4 antibody [EP618Y] (ab40759), expandable thumbnail

    Western blot - Anti-Smad4 antibody [EP618Y] (ab40759)

    Blocking and diluting buffer: 5% NFDM/TBST.

    All lanes: Western blot - Anti-Smad4 antibody [EP618Y] (AB40759) at 1/10000 dilution

    All lanes: NIH/3T3 (Mouse embyro fibroblast cell line) cell lysate at 20 µg

    Secondary

    All lanes: Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated at 1/1000 dilution

    Predicted band size: 60 kDa

    Observed band size: 60 kDa

  • Western blot - Anti-Smad4 antibody [EP618Y] (ab40759), expandable thumbnail

    Western blot - Anti-Smad4 antibody [EP618Y] (ab40759)

    Blocking and diluting buffer: 5% NFDM/TBST.

    All lanes: Western blot - Anti-Smad4 antibody [EP618Y] (AB40759) at 1/5000 dilution

    All lanes: PC-12 (Rat adrenal gland pheochromocytoma cell line) cell lysate at 10 µg

    Secondary

    All lanes: Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated at 1/1000 dilution

    Predicted band size: 60 kDa

    Observed band size: 60 kDa

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Smad4 antibody [EP618Y] (ab40759), expandable thumbnail
    This image is courtesy of an anonymous customer review.

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Smad4 antibody [EP618Y] (ab40759)

    Unpurified ab40759 staining Smad4 in rat femur tissue sections by Immunohistochemistry (IHC-P - paraformaldehyde-fixed, paraffin-embedded sections). Tissue was fixed with formaldehyde and blocked with 1% BSA for 20 minutes at 22°C; antigen retrieval was by heat mediation in a citrate buffer pH6.0. Samples were incubated with primary antibody (1/200 in blocking buffer) for 2 hours at 20°C. An undiluted HRP-conjugated goat anti-rabbit IgG polyclonal (1/250) was used as the secondary antibody.

  • Western blot - Anti-Smad4 antibody [EP618Y] (ab40759), expandable thumbnail

    Western blot - Anti-Smad4 antibody [EP618Y] (ab40759)

    All lanes: Western blot - Anti-Smad4 antibody [EP618Y] (AB40759) at 1/5000 dilution

    All lanes: SHSY5Y (Human neuroblastoma cell line from bone marrow) cell lysate at 10 µg

    Predicted band size: 60 kDa

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Smad4 antibody [EP618Y] (ab40759), expandable thumbnail

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Smad4 antibody [EP618Y] (ab40759)

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human breast carcinoma tissue labelling Smad4 with purified ab40759 at 1/100. Heat mediated antigen retrieval was performed using Tris/EDTA buffer pH 9. ab97051, a HRP-conjugated goat anti-rabbit IgG (H+L) was used as the secondary antibody (1/500). Negative control using PBS instead of primary antibody. Counterstained with hematoxylin.

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Product protocols

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

Please note: All products are 'FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC OR THERAPEUTIC PROCEDURES'.

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