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Rabbit Recombinant Monoclonal SMAD3 antibody. Suitable for IP, ChIP, WB and reacts with Human, Mouse, Rat, Recombinant fragment - Human samples. Cited in 15 publications.

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Images

ChIP - Anti-Smad3 antibody [EPR19686] - ChIP Grade (AB208182), expandable thumbnail
  • Western blot - Anti-Smad3 antibody [EPR19686] - ChIP Grade (AB208182), expandable thumbnail
  • Western blot - Anti-Smad3 antibody [EPR19686] - ChIP Grade (AB208182), expandable thumbnail
  • Immunoprecipitation - Anti-Smad3 antibody [EPR19686] - ChIP Grade (AB208182), expandable thumbnail
  • Western blot - Anti-Smad3 antibody [EPR19686] - ChIP Grade (AB208182), 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.

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Reactivity data

Select an application
Product promiseTestedExpectedPredictedNot recommended
IPChIPWB
Human
Tested
Tested
Tested
Mouse
Expected
Expected
Tested
Rat
Expected
Expected
Tested
Recombinant fragment - Human
Not recommended
Not recommended
Tested

Tested
Tested

Species
Human
Dilution info
1/30
Notes

-

Expected
Expected

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

-

Not recommended
Not recommended

Species
Recombinant fragment - Human
Dilution info
-
Notes

-

Tested
Tested

Species
Human
Dilution info
2 µg chromatin for 25 µg chromatin
Notes

-

Expected
Expected

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

-

Not recommended
Not recommended

Species
Recombinant fragment - Human
Dilution info
-
Notes

-

Tested
Tested

Species
Recombinant fragment - Human
Dilution info
1/1000
Notes

-

Species
Mouse
Dilution info
1/1000
Notes

-

Species
Rat
Dilution info
1/1000
Notes

-

Species
Human
Dilution info
1/1000
Notes

-

Associated Products

Select an associated product type

15 products for Alternative Product

Target data

Function

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.

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

Recommended products

Rabbit Recombinant Monoclonal SMAD3 antibody. Suitable for IP, ChIP, WB and reacts with Human, Mouse, Rat, Recombinant fragment - Human samples. Cited in 15 publications.

Key facts

Isotype
IgG
Form
Liquid
Clonality
Monoclonal
Immunogen
  • The exact immunogen used to generate this antibody is proprietary information.
Clone number
EPR19686
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

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.

Supplementary info

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

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.

Associated diseases and disorders

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.

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.

8 product images

  • ChIP - Anti-Smad3 antibody [EPR19686] - ChIP Grade (ab208182), expandable thumbnail

    ChIP - Anti-Smad3 antibody [EPR19686] - ChIP Grade (ab208182)

    Chromatin was prepared from HaCaT (Human keratinocyte cell line) cells treated with 7ng/ml TGF-β for 1h and non-treated according to the Abcam X-ChIP protocol. Cells were fixed with formaldehyde for 10 minutes. The ChIP was performed with 25μg of chromatin, 2μg of ab208182 (red), and 20μl of protein A/G beads. 2μg of rabbit normal IgG was added to the beads control (grey). The immunoprecipitated DNA was quantified by real time PCR (Sybr green approach).

    The ChIP condition performed here is similar to the literature (PMID: 18245174).

  • Western blot - Anti-Smad3 antibody [EPR19686] - ChIP Grade (ab208182), expandable thumbnail

    Western blot - Anti-Smad3 antibody [EPR19686] - ChIP Grade (ab208182)

    Lanes 1 - 4: Merged signal (red and green). Green - ab208182 observed at 50 kDa. Red - loading control, Anti-GAPDH antibody [6C5] - Loading Control ab8245 (Mouse anti-GAPDH antibody [6C5]) observed at 37kDa.

    ab208182 was shown to react with Smad3 in western blot. Membranes were blocked in 3% milk in TBS-T (0.1% Tween®) before incubation with ab208182 and Anti-GAPDH antibody [6C5] - Loading Control ab8245 (Mouse anti-GAPDH antibody [6C5]) overnight at 4°C at a 1 in 1000 Dilution and a 1 in 20000 dilution respectively. Blots were incubated 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 in 20000 dilution for 1 hour at room temperature before imaging.

    All lanes: Western blot - Anti-Smad3 antibody [EPR19686] - ChIP Grade (ab208182) at 1/1000 dilution

    Lane 1: Wild-type A549 (Human lung carcinoma cell line) whole cell lysate at 20 µg

    Lane 2: Western blot - Human SMAD3 knockout A549 cell lysate (Human SMAD3 knockout A549 cell lysate ab264513)

    Lane 3: HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate at 20 µg

    Lane 4: Human Kidney cell lysate at 20 µg

    Performed under reducing conditions.

    Predicted band size: 48 kDa

    Observed band size: 50 kDa

  • Western blot - Anti-Smad3 antibody [EPR19686] - ChIP Grade (ab208182), expandable thumbnail

    Western blot - Anti-Smad3 antibody [EPR19686] - ChIP Grade (ab208182)

    Lanes 1 - 4: Merged signal (red and green). Green - ab208182 observed at 55 kDa. Red - loading control, Anti-GAPDH antibody [6C5] - Loading Control ab8245 observed at 37 kDa.

    ab208182 was shown to react with Smad3 in wild-type HeLa cells. Loss of signal was observed when knockout cell line Human SMAD3 knockout HeLa cell line ab255431 (knockout cell lysate Human SMAD3 knockout HeLa cell lysate ab263834) was used. Wild-type and Smad3 knockout samples were subjected to SDS-PAGE. ab208182 and Anti-GAPDH antibody [6C5] - Loading Control (Anti-GAPDH antibody [6C5] - Loading Control ab8245) were incubated overnight at 4°C at 1 in 1000 dilution and 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.

    All lanes: Western blot - Anti-Smad3 antibody [EPR19686] - ChIP Grade (ab208182) at 1/1000 dilution

    Lane 1: Wild-type A549 cell lysate at 20 µg

    Lane 2: SMAD3 knockout A549 cell lysate at 20 µg

    Lane 3: Wild-type HeLa cell lysate at 20 µg

    Lane 4: Western blot - Human SMAD3 knockout HeLa cell lysate (Human SMAD3 knockout HeLa cell lysate ab263834)

    Secondary

    All lanes: Western blot - Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed ab216773) at 1/20000 dilution

    Performed under reducing conditions.

    Predicted band size: 48 kDa

    Observed band size: 37 kDa, 74 kDa

  • Immunoprecipitation - Anti-Smad3 antibody [EPR19686] - ChIP Grade (ab208182), expandable thumbnail

    Immunoprecipitation - Anti-Smad3 antibody [EPR19686] - ChIP Grade (ab208182)

    Smad3 was immunoprecipitated from 0.35 mg of HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate with ab208182 at 1/30 dilution.

    Western blot was performed from the immunoprecipitate using ab208182 at 1/1000 dilution.

    VeriBlot for IP Detection Reagent (HRP) (VeriBlot for IP Detection Reagent (HRP) ab131366), was used for detection at 1/10000 dilution.

    Lane 1: HeLa whole cell lysate 10μg (Input).

    Lane 2: ab208182 IP in HeLa whole cell lysate.

    Lane 3: Rabbit monoclonal IgG (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) instead of ab208182 in HeLa whole cell lysate.

    Blocking and dilution buffer and concentration: 5% NFDM/TBST.

    Exposure time: 3 minutes.

    All lanes: Immunoprecipitation - Anti-Smad3 antibody [EPR19686] - ChIP Grade (ab208182)

    Predicted band size: 48 kDa

  • Western blot - Anti-Smad3 antibody [EPR19686] - ChIP Grade (ab208182), expandable thumbnail

    Western blot - Anti-Smad3 antibody [EPR19686] - ChIP Grade (ab208182)

    Blocking/Dilution buffer: 5% NFDM/TBST.

    Human Smad3 recombinant protein contains aa2-227 with GST and His-Tag®. This protein was made-in house.

    All lanes: Western blot - Anti-Smad3 antibody [EPR19686] - ChIP Grade (ab208182) at 1/1000 dilution

    Lane 1: Human Smad3 recombinant protein at 0.01 µg

    Lane 2: Human Smad2 active protein at 0.01 µg

    Secondary

    All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/100000 dilution

    Predicted band size: 48 kDa

    Observed band size: 74 kDa

    Exposure time: 1s

  • Western blot - Anti-Smad3 antibody [EPR19686] - ChIP Grade (ab208182), expandable thumbnail

    Western blot - Anti-Smad3 antibody [EPR19686] - ChIP Grade (ab208182)

    Blocking/Dilution buffer: 5% NFDM/TBST.

    Exposure time: Lane 1/6: 3 minutes; Lane 2-5: 15 seconds.

    All lanes: Western blot - Anti-Smad3 antibody [EPR19686] - ChIP Grade (ab208182) at 1/1000 dilution

    Lane 1: HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate at 10 µg

    Lane 2: Jurkat (Human T cell leukemia cell line from peripheral blood) whole cell lysate at 10 µg

    Lane 3: HEK-293 (Human epithelial cell line from embryonic kidney) whole cell lysate at 10 µg

    Lane 4: K562 (Human chronic myelogenous leukemia cell line from bone marrow) whole cell lysate at 10 µg

    Lane 5: BxPC-3 (Human pancreas adenocarcinoma cell line) whole cell lysate at 10 µg

    Lane 6: Human fetal kidney lysate at 10 µg

    Secondary

    Lanes 1 - 5: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/100000 dilution

    Lane 6: Goat Anti-Rabbit IgG Peroxidase Conjugate, specific to the non-reduced form of IgG at 1/10000 dilution

    Predicted band size: 13 kDa, 48 kDa

    Observed band size: 55 kDa

  • Western blot - Anti-Smad3 antibody [EPR19686] - ChIP Grade (ab208182), expandable thumbnail

    Western blot - Anti-Smad3 antibody [EPR19686] - ChIP Grade (ab208182)

    Blocking/Dilution buffer: 5% NFDM/TBST.

    Exposure time: Lane 1/2: 30 seconds; Lane 3-6: 3 minutes.

    All lanes: Western blot - Anti-Smad3 antibody [EPR19686] - ChIP Grade (ab208182) at 1/1000 dilution

    Lane 1: C6 (Rat glial tumor cell line) whole cell lysate at 10 µg

    Lane 2: PC-12 (Rat adrenal gland pheochromocytoma cell line) whole cell lysate at 10 µg

    Lane 3: NIH/3T3 (Mouse embryonic fibroblast cell line) whole cell lysate at 10 µg

    Lane 4: Rat spleen lysate at 10 µg

    Lane 5: Rat kidney lysate at 10 µg

    Lane 6: Mouse spleen lysate at 10 µg

    Secondary

    All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/100000 dilution

    Predicted band size: 48 kDa

    Observed band size: 55 kDa

  • Western blot - Anti-Smad3 antibody [EPR19686] - ChIP Grade (ab208182), expandable thumbnail

    Western blot - Anti-Smad3 antibody [EPR19686] - ChIP Grade (ab208182)

    False colour image of Western blot: Anti-Smad3 antibody [EPR19686] - ChIP Grade staining at 1/1000 dilution, shown in green; Mouse anti-GAPDH antibody [6C5] (Anti-GAPDH antibody [6C5] - Loading Control ab8245) loading control staining at 1/20000 dilution, shown in red. In Western blot, ab208182 was shown to bind specifically to Smad3. A band was observed at 50 kDa in wild-type A549 cell lysates with no signal observed at this size in SMAD3 CRISPR-Cas9 edited cell line Human SMAD3 knockout A549 cell line ab277888 (CRISPR-Cas9 edited cell lysate None). The band observed in the CRISPR-Cas9 edited lysate lane below 50 kDa is likely to represent a truncated form of Smad3. This has not been investigated further and the functional properties of the gene product have not been determined. To generate this image, wild-type and SMAD3 CRISPR-Cas9 edited A549 cell lysates were analysed. First, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 3 % milk in TBS-0.1 % Tween® 20 (TBS-T) before incubation with primary antibodies overnight at 4 °C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged. Secondary antibodies used were Goat anti-Rabbit IgG H&L 800CW and Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution.

    All lanes: Western blot - Anti-Smad3 antibody [EPR19686] - ChIP Grade (ab208182) at 1/1000 dilution

    Lane 1: Wild-type A549 cell lysate at 20 µg

    Lane 2: SMAD3 CRISPR-Cas9 edited A549 cell lysate at 20 µg

    Secondary

    Lanes 1 - 2: Goat anti-Rabbit IgG H&L 800CW at 1/20000 dilution

    Lanes 1 - 2: Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution

    Performed under reducing conditions.

    Observed band size: 50 kDa

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