Name: Anti-Smad4 antibody [EP618Y]
SKU: ab40759
Rating: 4 (9 reviews)

Anti-Smad4 antibody [EP618Y] is a rabbit recombinant monoclonal antibody that is used to detect Smad4 in IHC-P, Western blot. Suitable for Human, Mouse, Rat samples.

- Specificity confirmed with SMAD4 knockout cell line validation
- Specificity and sensitivity confirmed in IHC with multi-tissue microarray (TMA) validation
- Cited in over 160 publications
- Trusted since 2006

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Anti-Smad4 antibody [EP618Y] - BSA and Azide free
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Alexa Fluor® 568 Anti-Smad4 antibody [EP618Y]
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Alexa Fluor® 555 Anti-Smad4 antibody [EP618Y]
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Alexa Fluor® 594 Anti-Smad4 antibody [EP618Y]
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Alexa Fluor® 647 Anti-Smad4 antibody [EP618Y]
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Alexa Fluor® 488 Anti-Smad4 antibody [EP618Y]
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Alexa Fluor® 750 Anti-Smad4 antibody [EP618Y]

Images

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Smad4 antibody [EP618Y] (AB40759), expandable thumbnail

Publications

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.

Abcam Recommends

Reactivity data

Select an application

Product promiseTestedExpectedPredictedNot recommended

	IHC-P	IP	Flow Cyt	WB	ICC/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	Dilution info	Notes
Species Human	Dilution info 1/100	Notes Perform heat-mediated antigen retrieval before commencing with IHC staining protocol.

Expected
Expected

Species	Dilution info	Notes
Species Mouse, Rat	Dilution info Use at an assay dependent concentration.	Notes -

Not recommended
Not recommended

Species	Dilution info	Notes
Species Mouse, Rat, Human	Dilution info -	Notes -

Not recommended
Not recommended

Species	Dilution info	Notes
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	Dilution info	Notes
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	Dilution info	Notes
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.

Associated Products

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6 products for Alternative Product

Target data

See full target information SMAD4

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

DPC4, MADH4, SMAD4, Mothers against decapentaplegic homolog 4, MAD homolog 4, Mothers against DPP homolog 4, Deletion target in pancreatic carcinoma 4, SMAD family member 4, SMAD 4, Smad4, hSMAD4

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.
Clone number: EP618Y
Purification technique: Affinity purification Protein A
Concentration

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

Product Specifications
Anti-Smad4 antibody [EP618Y] (ab40759) was developed by Abcam using patented rabbit monoclonal antibody technology and is validated for use in IHC-P, WB in human, mouse, rat samples.
Anti-Smad4 antibody [EP618Y] (ab40759) specifically detects Smad4 (UniProt ID: Q13485; Molecular weight: 60kDa) and is sold in 100 µL and 1 mL selling sizes.

Quality and Validation
Abcam's high quality manufacturing and validation processes ensure Anti-Smad4 antibody [EP618Y] (ab40759) has high sensitivity and specificity alongside high lot-to-lot consistency and reproducibility.
The specificity of Anti-Smad4 antibody [EP618Y] (ab40759) has been confirmed by testing in knockout samples.
Anti-Smad4 antibody [EP618Y] (ab40759) has been cited over 170 times in peer reviewed journals and is trusted by the scientific community.
Anti-Smad4 antibody [EP618Y] (ab40759) has 9 independent reviews from customers.

Related Products
Antibody clone EP618Y is also available pre-conjugated to a variety of labels for your convenience - HRP, Alexa Fluor® 488, Alexa Fluor® 647, Alexa Fluor® 594, Alexa Fluor® 568, Alexa Fluor® 555, Alexa Fluor® 750 (ab195554, ab311050, ab311176, ab311831, ab313114, ab313307, ab321616).

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

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. Goat Anti-Rabbit IgG H&L (HRP) 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.
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, Anti-GAPDH antibody [mAbcam 9484] - 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 Anti-GAPDH antibody [mAbcam 9484] - Loading Control 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 Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed ab216773 and Goat anti-Mouse IgG H&L (IRDye^® 680RD) preabsorbed Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed 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)
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)
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)
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) (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)
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) (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)
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)
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)
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
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)
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