Rabbit Recombinant Monoclonal SMAD3 antibody. Carrier free. Suitable for ChIC/CUT&RUN-seq, WB, IP, ChIP, ICC/IF, Flow Cyt (Intra) and reacts with Human, Recombinant fragment - Human samples. Cited in 1 publication.
Images
![Immunoprecipitation - Anti-Smad2 + Smad3 antibody [EPR19557] - BSA and Azide free (AB236030), expandable thumbnail](https://content.abcam.com/products/images/smad2-smad3-antibody-epr19557-bsa-and-azide-free-ab236030--immunoprecipitation-img24792.jpg/_jcr_content/renditions/webp-475.webp "Immunoprecipitation - Anti-Smad2 + Smad3 antibody [EPR19557] - BSA and Azide free (AB236030)")
![ChIP - Anti-Smad2 + Smad3 antibody [EPR19557] - BSA and Azide free (AB236030), expandable thumbnail](https://content.abcam.com/products/images/smad2-smad3-antibody-epr19557-bsa-and-azide-free-ab236030--chip-img13118.jpg/_jcr_content/renditions/webp-475.webp "ChIP - Anti-Smad2 + Smad3 antibody [EPR19557] - BSA and Azide free (AB236030)")
![Immunocytochemistry/ Immunofluorescence - Anti-Smad2 + Smad3 antibody [EPR19557] - BSA and Azide free (AB236030), expandable thumbnail](https://content.abcam.com/products/images/smad2-smad3-antibody-epr19557-bsa-and-azide-free-ab236030--immunocytochemistry-immunofluorescence-img64255.jpg/_jcr_content/renditions/webp-475.webp "Immunocytochemistry/ Immunofluorescence - Anti-Smad2 + Smad3 antibody [EPR19557] - BSA and Azide free (AB236030)")
![Immunoprecipitation - Anti-Smad2 + Smad3 antibody [EPR19557] - BSA and Azide free (AB236030), expandable thumbnail](https://content.abcam.com/products/images/smad2-smad3-antibody-epr19557-bsa-and-azide-free-ab236030--immunoprecipitation-img3804.jpg/_jcr_content/renditions/webp-475.webp "Immunoprecipitation - Anti-Smad2 + Smad3 antibody [EPR19557] - BSA and Azide free (AB236030)")
![Flow Cytometry (Intracellular) - Anti-Smad2 + Smad3 antibody [EPR19557] - BSA and Azide free (AB236030), expandable thumbnail](https://content.abcam.com/products/images/smad2-smad3-antibody-epr19557-bsa-and-azide-free-ab236030--flow-cytometry-intracellular-img6145.jpg/_jcr_content/renditions/webp-475.webp "Flow Cytometry (Intracellular) - Anti-Smad2 + Smad3 antibody [EPR19557] - BSA and Azide free (AB236030)")
Publications
Filter by
- Medical science monitor : international medical jo 25:3069-30762019Mesenchymal Stem Cells Reverse Diabetic Nephropathy Disease via Lipoxin A4 by Targeting Transforming Growth Factor β (TGF-β)/smad Pathway and Pro-Inflammatory Cytokines.Applications:Unspecified applicationReactive species:Unspecified reactive speciesYihua Bai et. al.PubMed 31023998
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| ChIC/CUT&RUN-seq | WB | IP | ChIP | ICC/IF | Flow Cyt (Intra) | |
|---|---|---|---|---|---|---|
| Human | Tested | Expected | Tested | Tested | Tested | Tested |
| Recombinant fragment - Human | Not recommended | Tested | Not recommended | Not recommended | Not recommended | Not recommended |
ChIC/CUT&RUN-seq
TestedTested
| Species | Dilution info | Notes |
|---|---|---|
Species Human | Dilution info - | Notes - |
Not recommendedNot recommended
| Species | Dilution info | Notes |
|---|---|---|
Species Recombinant fragment - Human | Dilution info - | Notes - |
WB
TestedTested
| Species | Dilution info | Notes |
|---|---|---|
Species Recombinant fragment - Human | Dilution info - | Notes - |
ExpectedExpected
| Species | Dilution info | Notes |
|---|---|---|
Species Human | Dilution info Use at an assay dependent concentration. | Notes - |
IP
TestedTested
| Species | Dilution info | Notes |
|---|---|---|
Species Human | Dilution info - | Notes - |
Not recommendedNot recommended
| Species | Dilution info | Notes |
|---|---|---|
Species Recombinant fragment - Human | Dilution info - | Notes - |
ChIP
TestedTested
| Species | Dilution info | Notes |
|---|---|---|
Species Human | Dilution info - | Notes - |
Not recommendedNot recommended
| Species | Dilution info | Notes |
|---|---|---|
Species Recombinant fragment - Human | Dilution info - | Notes - |
ICC/IF
TestedTested
| Species | Dilution info | Notes |
|---|---|---|
Species Human | Dilution info - | Notes - |
Not recommendedNot recommended
| Species | Dilution info | Notes |
|---|---|---|
Species Recombinant fragment - Human | Dilution info - | Notes - |
Flow Cyt (Intra)
TestedTested
| Species | Dilution info | Notes |
|---|---|---|
Species Human | Dilution info - | Notes - |
Not recommendedNot recommended
| Species | Dilution info | Notes |
|---|---|---|
Species Recombinant fragment - Human | Dilution info - | Notes - |
Associated Products
Select an associated product type
4 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.
Additional Targets
SMAD2
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 ChIC/CUT&RUN-seq, WB, IP, ChIP, ICC/IF, Flow Cyt (Intra) and reacts with Human, Recombinant fragment - Human samples. Cited in 1 publication.
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
- EPR19557
- 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
ab236030 is the carrier-free version of Anti-Smad2 + Smad3 antibody [EPR19557] - ChIP Grade ab207447.
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
Smad2 and Smad3 are intracellular proteins tightly involved in the TGF-β signaling pathway. These proteins also known as mothers against decapentaplegic homolog 2 and 3 play an important role in transmitting signals from the cell surface to the nucleus. Smad2 has a molecular weight of approximately 58 kDa while Smad3 is about 52 kDa. They are expressed in various tissues throughout the body including heart skin and immune cells. Upon activation Smad2 and Smad3 undergo phosphorylation a modification that is important for their function as signal transducers.
- Biological function summary
The functions of Smad2 and Smad3 are critical in mediating cellular processes such as proliferation differentiation and apoptosis. These proteins often form complexes specifically oligomeric structures with other Smad proteins like co-Smad4 to exert their regulatory roles. The Smad2/3 complexes control the transcription of specific genes in response to TGF-β stimuli influencing cellular responses in a context-dependent manner.
- Pathways
Smad2 and Smad3 are central to the TGF-β signaling pathway one significant pathway that affects cellular growth and differentiation. These proteins interact with TGF-β receptors on the cell surface to become phosphorylated forming a complex with Smad4 which then translocates to the nucleus. In the nucleus this complex regulates gene expression. Both Smad2 and Smad3 also engage with other signaling pathways such as the MAPK pathway further influencing cell behavior and cooperating with proteins like ERK.
- Associated diseases and disorders
Smad2 and Smad3 have been linked to conditions like cancer and fibrosis. Dysregulation of Smad2/3 signaling often leads to unchecked cell proliferation a hallmark of tumorigenesis. In fibrosis excessive Smad2/3 activity contributes to tissue scarring due to increased extracellular matrix production. For instance their interaction with proteins like α-SMA (alpha-smooth muscle actin) in fibroblasts plays a role in cardiac and pulmonary fibrosis. Understanding their exact contribution provides insights into potential therapeutic targets for treating these diseases.
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.
6 product images
![Immunoprecipitation - Anti-Smad2 + Smad3 antibody [EPR19557] - BSA and Azide free (ab236030), expandable thumbnail](https://content.abcam.com/products/images/smad2-smad3-antibody-epr19557-bsa-and-azide-free-ab236030--immunoprecipitation-img24792.jpg "Immunoprecipitation - Anti-Smad2 + Smad3 antibody [EPR19557] - BSA and Azide free (ab236030)")
Immunoprecipitation - Anti-Smad2 + Smad3 antibody [EPR19557] - BSA and Azide free (ab236030)
Smad2 + Smad3 were immunoprecipitated from 1mg of HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate with Anti-Smad2 + Smad3 antibody [EPR19557] - ChIP Grade ab207447 at 1/40 dilution. Western blot was performed from the immunoprecipitate using Anti-Smad2 + Smad3 antibody [EPR19557] - ChIP Grade ab207447 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: Anti-Smad2 + Smad3 antibody [EPR19557] - ChIP Grade ab207447 IP in HeLa whole cell lysate.
Lane 3: Rabbit monoclonal IgG (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) instead of Anti-Smad2 + Smad3 antibody [EPR19557] - ChIP Grade ab207447 in HeLa whole cell lysate.
Blocking and dilution buffer and concentration: 5% NFDM/TBST.
Exposure time: 5 seconds.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-Smad2 + Smad3 antibody [EPR19557] - ChIP Grade ab207447).
All lanes: Immunoprecipitation - Anti-Smad2 + Smad3 antibody [EPR19557] - ChIP Grade (Anti-Smad2 + Smad3 antibody [EPR19557] - ChIP Grade ab207447)
![ChIP - Anti-Smad2 + Smad3 antibody [EPR19557] - BSA and Azide free (ab236030), expandable thumbnail](https://content.abcam.com/products/images/smad2-smad3-antibody-epr19557-bsa-and-azide-free-ab236030--chip-img13118.jpg "ChIP - Anti-Smad2 + Smad3 antibody [EPR19557] - BSA and Azide free (ab236030)")
ChIP - Anti-Smad2 + Smad3 antibody [EPR19557] - BSA and Azide free (ab236030)
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-Smad2 + Smad3 antibody [EPR19557] - ChIP Grade ab207447 (blue), and 20µl of Anti rabit IgG sepharose beads. 2μg of rabbit normal IgG was added to the beads control (yellow). The immunoprecipitated DNA was quantified by real time PCR (Sybr green approach).
The ChIP condition is designed against Smad2 refer to PMID: 18955504.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-Smad2 + Smad3 antibody [EPR19557] - ChIP Grade ab207447).
![Immunocytochemistry/ Immunofluorescence - Anti-Smad2 + Smad3 antibody [EPR19557] - BSA and Azide free (ab236030), expandable thumbnail](https://content.abcam.com/products/images/smad2-smad3-antibody-epr19557-bsa-and-azide-free-ab236030--immunocytochemistry-immunofluorescence-img64255.jpg "Immunocytochemistry/ Immunofluorescence - Anti-Smad2 + Smad3 antibody [EPR19557] - BSA and Azide free (ab236030)")
Immunocytochemistry/ Immunofluorescence - Anti-Smad2 + Smad3 antibody [EPR19557] - BSA and Azide free (ab236030)
Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized HeLa (Human epithelial cell line from cervix adenocarcinoma) cells labeling Smad2 + Smad3 with Anti-Smad2 + Smad3 antibody [EPR19557] - ChIP Grade ab207447 at 1/500 dilution, followed by Goat Anti-Rabbit IgG (Alexa Fluor® 488) (Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077) secondary antibody at 1/1000 dilution (green). Confocal image showing signal translocation from cytoplasm to nucleus after TGF-beta (10ng/ml, 1h) treatment in HeLa cells. PMID: 9006934. The nuclear counter stain is DAPI (blue).
Tubulin is detected with Anti-alpha Tubulin mouse MAb (Anti-alpha Tubulin antibody [DM1A] - Loading Control ab7291) at 1/1000 dilution, followed by Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) (Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) preadsorbed ab150120) secondary antibody at 1/1000 dilution (red).
The negative controls are as follows:
-ve control 1: Anti-Smad2 + Smad3 antibody [EPR19557] - ChIP Grade ab207447 at 1/500 dilution, followed by Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) (Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) preadsorbed ab150120) secondary at 1/1000 dilution.
-ve control 2: Anti-alpha Tubulin mouse MAb (Anti-alpha Tubulin antibody [DM1A] - Loading Control ab7291) at 1/1000 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077) secondary at 1/1000 dilution.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-Smad2 + Smad3 antibody [EPR19557] - ChIP Grade ab207447).
![Immunoprecipitation - Anti-Smad2 + Smad3 antibody [EPR19557] - BSA and Azide free (ab236030), expandable thumbnail](https://content.abcam.com/products/images/smad2-smad3-antibody-epr19557-bsa-and-azide-free-ab236030--immunoprecipitation-img3804.jpg "Immunoprecipitation - Anti-Smad2 + Smad3 antibody [EPR19557] - BSA and Azide free (ab236030)")
Immunoprecipitation - Anti-Smad2 + Smad3 antibody [EPR19557] - BSA and Azide free (ab236030)
Anti-Smad2 + Smad3 antibody [EPR19557] - ChIP Grade ab207447 Immunoprecipitating Smad2 + Smad3 in human HeLa whole cell lysate . 2μg of capture antibody in 0.35mg lysate was used. 10μg of cell lysate was incubated with primary antibody at a dilution of 1/1000 and VeriBlot for IP Detection Reagent (HRP) VeriBlot for IP Detection Reagent (HRP) ab131366 was used for detection at a dilution of 1/1000.
Lane 1: HeLa (human cervix adenocarcinoma) whole cell lysate
Lane 2: HeLa (human cervix adenocarcinoma) whole cell lysate
Lane 3: Rabbit monoclonal IgG (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) instead of Anti-Smad2 + Smad3 antibody [EPR19557] - ChIP Grade ab207447 in HeLa (human cervix adenocarcinoma) whole cell lysateBlocking buffer: 5% NFDM/TBST
Diluting buffer: 5% NFDM/TBST
Smad2 and Smad3 can be resolved using a lower percentage gel.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-Smad2 + Smad3 antibody [EPR19557] - ChIP Grade ab207447).
All lanes: Immunoprecipitation - Anti-Smad2 + Smad3 antibody [EPR19557] - ChIP Grade (Anti-Smad2 + Smad3 antibody [EPR19557] - ChIP Grade ab207447)
Observed band size: 58-62 kDa
Exposure time: 3min
![Flow Cytometry (Intracellular) - Anti-Smad2 + Smad3 antibody [EPR19557] - BSA and Azide free (ab236030), expandable thumbnail](https://content.abcam.com/products/images/smad2-smad3-antibody-epr19557-bsa-and-azide-free-ab236030--flow-cytometry-intracellular-img6145.jpg "Flow Cytometry (Intracellular) - Anti-Smad2 + Smad3 antibody [EPR19557] - BSA and Azide free (ab236030)")
Flow Cytometry (Intracellular) - Anti-Smad2 + Smad3 antibody [EPR19557] - BSA and Azide free (ab236030)
Intracellular flow cytometric analysis of 4% paraformaldehyde-fixed HeLa (Human epithelial cell line from cervix adenocarcinoma) cells labeling Smad2 and Smad3 with Anti-Smad2 + Smad3 antibody [EPR19557] - ChIP Grade ab207447 at 1/500 dilution (red) compared with a rabbit monoclonal IgG isotype control (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730; black) and an unlabelled control (cells without incubation with primary antibody and secondary antibody; blue). Goat anti rabbit IgG (Alexa Fluor® 488) at 1/500 dilution was used as the secondary antibody.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-Smad2 + Smad3 antibody [EPR19557] - ChIP Grade ab207447).
![ChIC/CUT&RUN sequencing - Anti-Smad2 + Smad3 antibody [EPR19557] - BSA and Azide free (ab236030), expandable thumbnail](https://content.abcam.com/products/images/smad2-smad3-antibody-epr19557-bsa-and-azide-free-ab236030--chic-cut-run-sequencing-img419269.jpg "ChIC/CUT&RUN sequencing - Anti-Smad2 + Smad3 antibody [EPR19557] - BSA and Azide free (ab236030)")
ChIC/CUT&RUN sequencing - Anti-Smad2 + Smad3 antibody [EPR19557] - BSA and Azide free (ab236030)
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-Smad2 + Smad3 antibody [EPR19557] - ChIP Grade ab207447).
ChIC/CUT&RUN was performed using a pAG-MNAse at a final concentration of 700 ng/mL, 2.5 x 10^5 HaCaT (Human keratinocyte cell line) cells (treated with 7ng/ml TGF-β for 1h) and 5 µg of Anti-Smad2 + Smad3 antibody [EPR19557] - ChIP Grade ab207447 [EPR19557]. The resulting DNA was sequenced on the Illumina NovaSeq 6000 to a depth of 10 million reads. The negative IgG control Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730 is also shown.
Additional screenshots of mapped reads can be found in the Protocol booklet in the Support and downloads section. The University of Geneva owns patents relevant to ChIC (Chromatin Immuno-Cleavage) methods.
![ChIC/CUT&RUN sequencing - Anti-Smad2 + Smad3 antibody [EPR19557] - BSA and Azide free (ab236030), expandable thumbnail](https://content.abcam.com/products/images/smad2-smad3-antibody-epr19557-bsa-and-azide-free-ab236030--chic-cut-run-sequencing-img419269.jpg/_jcr_content/renditions/webp-475.webp "ChIC/CUT&RUN sequencing - Anti-Smad2 + Smad3 antibody [EPR19557] - BSA and Azide free (ab236030)")
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