Rabbit Recombinant Monoclonal SMAD3 antibody. Carrier free. Suitable for IP, ChIP, WB and reacts with Recombinant fragment, Mouse, Rat, Human samples.
Images
![ChIP - Anti-Smad3 antibody [EPR19686] - BSA and Azide free (AB251490), expandable thumbnail](https://content.abcam.com/products/images/smad3-antibody-epr19686-bsa-and-azide-free-ab251490--chip-img99819.jpg/_jcr_content/renditions/webp-475.webp "ChIP - Anti-Smad3 antibody [EPR19686] - BSA and Azide free (AB251490)")
![Western blot - Anti-Smad3 antibody [EPR19686] - BSA and Azide free (AB251490), expandable thumbnail](https://content.abcam.com/products/images/smad3-antibody-epr19686-bsa-and-azide-free-ab251490--western-blot-img140534.jpg/_jcr_content/renditions/webp-475.webp "Western blot - Anti-Smad3 antibody [EPR19686] - BSA and Azide free (AB251490)")
![Western blot - Anti-Smad3 antibody [EPR19686] - BSA and Azide free (AB251490), expandable thumbnail](https://content.abcam.com/products/images/smad3-antibody-epr19686-bsa-and-azide-free-ab251490--western-blot-img104837.jpg/_jcr_content/renditions/webp-475.webp "Western blot - Anti-Smad3 antibody [EPR19686] - BSA and Azide free (AB251490)")
![Western blot - Anti-Smad3 antibody [EPR19686] - BSA and Azide free (AB251490), expandable thumbnail](https://content.abcam.com/products/images/smad3-antibody-epr19686-bsa-and-azide-free-ab251490--western-blot-img418177.jpg/_jcr_content/renditions/webp-475.webp "Western blot - Anti-Smad3 antibody [EPR19686] - BSA and Azide free (AB251490)")
Key facts
- Isotype
- IgG
- Host species
- Rabbit
- Storage buffer
pH: 7.2 - 7.4
Constituents: PBS- Form
- Liquid
- Clonality
- Monoclonal
Immunogen
- The exact immunogen used to generate this antibody is proprietary information.
Reactivity data
Select an application| IP | ChIP | WB | |
|---|---|---|---|
| Human | Expected | Tested | Tested |
| Mouse | Expected | Predicted | Predicted |
| Rat | Expected | Predicted | Predicted |
| Recombinant fragment | Expected | Not recommended | Not recommended |
IP
ExpectedExpected
| Species | Dilution info | Notes |
|---|---|---|
Species Recombinant fragment, Mouse, Rat, Human | Dilution info Use at an assay dependent concentration. | Notes - |
ChIP
TestedTested
| Species | Dilution info | Notes |
|---|---|---|
Species Human | Dilution info 2 µg chromatin for 25.00000 µg chromatin | Notes - |
PredictedPredicted
| Species | Dilution info | Notes |
|---|---|---|
Species Mouse, Rat | Dilution info - | Notes - |
Not recommendedNot recommended
| Species | Dilution info | Notes |
|---|---|---|
Species Recombinant fragment | Dilution info - | Notes - |
WB
TestedTested
| Species | Dilution info | Notes |
|---|---|---|
Species Human | Dilution info - | Notes - |
PredictedPredicted
| Species | Dilution info | Notes |
|---|---|---|
Species Mouse, Rat | Dilution info - | Notes - |
Not recommendedNot recommended
| Species | Dilution info | Notes |
|---|---|---|
Species Recombinant fragment | Dilution info - | Notes - |
Target data
Function
Receptor-regulated SMAD (R-SMAD) that is an intracellular signal transducer and transcriptional modulator activated by TGF-beta (transforming growth factor) and activin type 1 receptor kinases. Binds the TRE element in the promoter region of many genes that are regulated by TGF-beta and, on formation of the SMAD3/SMAD4 complex, activates transcription. Also can form a SMAD3/SMAD4/JUN/FOS complex at the AP-1/SMAD site to regulate TGF-beta-mediated transcription. Has an inhibitory effect on wound healing probably by modulating both growth and migration of primary keratinocytes and by altering the TGF-mediated chemotaxis of monocytes. This effect on wound healing appears to be hormone-sensitive. Regulator of chondrogenesis and osteogenesis and inhibits early healing of bone fractures. Positively regulates PDPK1 kinase activity by stimulating its dissociation from the 14-3-3 protein YWHAQ which acts as a negative regulator.
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. Carrier free. Suitable for IP, ChIP, WB and reacts with Recombinant fragment, Mouse, Rat, Human samples.
Key facts
- Isotype
- IgG
- Host species
- Rabbit
- Storage buffer
pH: 7.2 - 7.4
Constituents: PBS- Form
- Liquid
- Clonality
- Monoclonal
- Immunogen
- The exact immunogen used to generate this antibody is proprietary information.
- Carrier free
- Yes
- Clone number
- EPR19686
- Purification technique
- Affinity purification Protein A
- Concentration
Storage
- Shipped at conditions
- Blue Ice
- Appropriate short-term storage conditions
- +4°C
- Appropriate long-term storage conditions
- +4°C
- Storage information
- Do Not Freeze
Notes
ab251490 is the carrier-free version of Anti-Smad3 antibody [EPR19686] - ChIP Grade ab208182.
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.
Our carrier-free antibodies are typically supplied in a PBS-only formulation, purified and free of BSA, sodium azide and glycerol. The carrier-free buffer and high concentration allow for increased conjugation efficiency.
This conjugation-ready format is designed for use with fluorochromes, metal isotopes, oligonucleotides, and enzymes, which makes them ideal for antibody labelling, functional and cell-based assays, flow-based assays (e.g. mass cytometry) and Multiplex Imaging applications.
Use our conjugation kits for antibody conjugates that are ready-to-use in as little as 20 minutes with 1 minute hands-on-time and 100% antibody recovery: available for fluorescent dyes, HRP, biotin and gold.
This product is compatible with the Maxpar® Antibody Labeling Kit from Fluidigm, without the need for antibody preparation. Maxpar® is a trademark of Fluidigm Canada Inc.
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
Tested
We have tested this species and application combination and it works. It is covered by our product promise.
Expected
We have not tested this specific species and application combination in-house, but expect it will work. It is covered by our product promise.
Predicted
This species and application combination has not been tested, but we predict it will work based on strong homology. However, this combination is not covered by our product promise.
Not recommended
We do not recommend this combination. It is not covered by our 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.
4 product images
![ChIP - Anti-Smad3 antibody [EPR19686] - BSA and Azide free (ab251490), expandable thumbnail](https://content.abcam.com/products/images/smad3-antibody-epr19686-bsa-and-azide-free-ab251490--chip-img99819.jpg "ChIP - Anti-Smad3 antibody [EPR19686] - BSA and Azide free (ab251490)")
ChIP - Anti-Smad3 antibody [EPR19686] - BSA and Azide free (ab251490)
This data was developed using the same antibody clone in a different buffer formulation (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 Anti-Smad3 antibody [EPR19686] - ChIP Grade 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] - BSA and Azide free (ab251490), expandable thumbnail](https://content.abcam.com/products/images/smad3-antibody-epr19686-bsa-and-azide-free-ab251490--western-blot-img140534.jpg "Western blot - Anti-Smad3 antibody [EPR19686] - BSA and Azide free (ab251490)")
Western blot - Anti-Smad3 antibody [EPR19686] - BSA and Azide free (ab251490)
This data was developed using the same antibody clone in a different buffer formulation (Anti-Smad3 antibody [EPR19686] - ChIP Grade ab208182).
Lanes 1 - 4: Merged signal (red and green). Green - Anti-Smad3 antibody [EPR19686] - ChIP Grade ab208182 observed at 50 kDa. Red - loading control, Anti-GAPDH antibody [6C5] - Loading Control ab8245 (Mouse anti-GAPDH antibody [6C5]) observed at 37kDa.
Anti-Smad3 antibody [EPR19686] - ChIP Grade 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 Anti-Smad3 antibody [EPR19686] - ChIP Grade 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 (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] - BSA and Azide free (ab251490), expandable thumbnail](https://content.abcam.com/products/images/smad3-antibody-epr19686-bsa-and-azide-free-ab251490--western-blot-img104837.jpg "Western blot - Anti-Smad3 antibody [EPR19686] - BSA and Azide free (ab251490)")
Western blot - Anti-Smad3 antibody [EPR19686] - BSA and Azide free (ab251490)
This data was developed using the same antibody clone in a different buffer formulation (Anti-Smad3 antibody [EPR19686] - ChIP Grade ab208182).
Lanes 1-4: Merged signal (red and green). Green - Anti-Smad3 antibody [EPR19686] - ChIP Grade ab208182 observed at 74 kDa. Red - loading control, Anti-GAPDH antibody [6C5] - Loading Control ab8245 observed at 37 kDa.
Anti-Smad3 antibody [EPR19686] - ChIP Grade ab208182 was shown to react with Smad3 in wildtype HeLa. Loss of signal was observed when knockout HeLa cell line Human SMAD3 knockout HeLa cell line ab255431 (knockout cell lysate Human SMAD3 knockout HeLa cell lysate ab263834). Wild-type and Smad3 knockout samples were subjected to SDS-PAGE. Anti-Smad3 antibody [EPR19686] - ChIP Grade 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 (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)
SecondaryAll 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
![Western blot - Anti-Smad3 antibody [EPR19686] - BSA and Azide free (ab251490), expandable thumbnail](https://content.abcam.com/products/images/smad3-antibody-epr19686-bsa-and-azide-free-ab251490--western-blot-img418177.jpg "Western blot - Anti-Smad3 antibody [EPR19686] - BSA and Azide free (ab251490)")
Western blot - Anti-Smad3 antibody [EPR19686] - BSA and Azide free (ab251490)
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, Anti-Smad3 antibody [EPR19686] - ChIP Grade 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 (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
SecondaryLanes 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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