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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.

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Images

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.
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Reactivity data

Select an application
Product promiseTestedExpectedPredictedNot recommended
ChIC/CUT&RUN-seqWBIPChIPICC/IFFlow Cyt (Intra)
Human
Tested
Expected
Tested
Tested
Tested
Tested
Recombinant fragment - Human
Not recommended
Tested
Not recommended
Not recommended
Not recommended
Not recommended

Tested
Tested

Species

Human

Dilution info

-

Notes

-

Not recommended
Not recommended

Species

Recombinant fragment - Human

Dilution info

-

Notes

-

Tested
Tested

Species

Recombinant fragment - Human

Dilution info

-

Notes

-

Expected
Expected

Species

Human

Dilution info

Use at an assay dependent concentration.

Notes

-

Tested
Tested

Species

Human

Dilution info

-

Notes

-

Not recommended
Not recommended

Species

Recombinant fragment - Human

Dilution info

-

Notes

-

Tested
Tested

Species

Human

Dilution info

-

Notes

-

Not recommended
Not recommended

Species

Recombinant fragment - Human

Dilution info

-

Notes

-

Tested
Tested

Species

Human

Dilution info

-

Notes

-

Not recommended
Not recommended

Species

Recombinant fragment - Human

Dilution info

-

Notes

-

Tested
Tested

Species

Human

Dilution info

-

Notes

-

Not recommended
Not recommended

Species

Recombinant fragment - Human

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

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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.

Alternative names

Key facts

Isotype

IgG

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
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Storage

Shipped at conditions

Blue Ice

Appropriate long-term storage conditions

+4°C

Storage information

Do Not Freeze

Notes

ab236030 is the carrier-free version of 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

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.

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.

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

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

    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 ab207447 at 1/40 dilution. Western blot was performed from the immunoprecipitate using ab207447 at 1/1000 dilution. 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: ab207447 IP in HeLa whole cell lysate.

    Lane 3: Rabbit monoclonal IgG (ab172730) instead of 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 (ab207447).

    All lanes: Immunoprecipitation - Anti-Smad2 + Smad3 antibody [EPR19557] - ChIP Grade (AB207447)

  • ChIP - Anti-Smad2 + Smad3 antibody [EPR19557] - BSA and Azide free (ab236030), expandable thumbnail

    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 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 (ab207447).

  • Immunocytochemistry/ Immunofluorescence - Anti-Smad2 + Smad3 antibody [EPR19557] - BSA and Azide free (ab236030), expandable thumbnail

    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 ab207447 at 1/500 dilution, followed by Goat Anti-Rabbit IgG (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 (ab7291) at 1/1000 dilution, followed by Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) (ab150120) secondary antibody at 1/1000 dilution (red).

    The negative controls are as follows:

    -ve control 1: ab207447 at 1/500 dilution, followed by Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) (ab150120) secondary at 1/1000 dilution.

    -ve control 2: Anti-alpha Tubulin mouse MAb (ab7291) at 1/1000 dilution, followed by 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 (ab207447).

  • Immunoprecipitation - Anti-Smad2 + Smad3 antibody [EPR19557] - BSA and Azide free (ab236030), expandable thumbnail

    Immunoprecipitation - Anti-Smad2 + Smad3 antibody [EPR19557] - BSA and Azide free (ab236030)

    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) 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 (ab172730) instead of ab207447 in HeLa (human cervix adenocarcinoma) whole cell lysate

    Blocking 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 (ab207447).

    All lanes: Immunoprecipitation - 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

    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 ab207447 at 1/500 dilution (red) compared with a rabbit monoclonal IgG 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 (ab207447).

  • ChIC/CUT&RUN sequencing - Anti-Smad2 + Smad3 antibody [EPR19557] - BSA and Azide free (ab236030), expandable thumbnail

    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 (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 ab207447 [EPR19557]. The resulting DNA was sequenced on the Illumina NovaSeq 6000 to a depth of 10 million reads. The negative IgG 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.

Downloads

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'.

For licensing inquiries, please contact partnerships@abcam.com

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