Name: Anti-Smad2 antibody [EP784Y]
SKU: ab40855
Rating: 5 (4 reviews)

Anti-Smad2 antibody [EP784Y] (ab40855) is a rabbit monoclonal antibody detecting Smad2 in Western Blot, Flow Cytometry (Intra), Flow Cytometry, IP, IHC-P, ICC/IF. Suitable for Human, Rat.

- KO validated for confirmed specificity

- Biophysical QC for unrivalled batch-batch consistency

- Over 100 publications

- Trusted since 2006

Images

Immunoprecipitation - Anti-Smad2 antibody [EP784Y] (AB40855), 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

	ChIC/CUT&RUN-seq	IP	WB	ICC/IF	Flow Cyt (Intra)	IHC-P
Human	Tested	Tested	Tested	Tested	Tested	Tested
Rat	Expected	Expected	Tested	Expected	Expected	Expected

Tested
Tested

Species	Dilution info	Notes
Species Human	Dilution info -	Notes -

Expected
Expected

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

Tested
Tested

Species	Dilution info	Notes
Species Human	Dilution info 1/20 - 1/50	Notes Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

Expected
Expected

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

Tested
Tested

Species	Dilution info	Notes
Species Rat	Dilution info 1/2000 - 1/10000	Notes -
Species Human	Dilution info 1/2000 - 1/10000	Notes -

Tested
Tested

Species	Dilution info	Notes
Species Human	Dilution info 1/100 - 1/250	Notes -

Expected
Expected

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

Tested
Tested

Species	Dilution info	Notes
Species Human	Dilution info 1/20 - 1/100	Notes Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.

Expected
Expected

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

Tested
Tested

Species	Dilution info	Notes
Species Human	Dilution info 1/50	Notes Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

Expected
Expected

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

Associated Products

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Target data

See full target information SMAD2

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 SMAD2/SMAD4 complex, activates transcription. Promotes TGFB1-mediated transcription of odontoblastic differentiation genes in dental papilla cells (By similarity). Positively regulates PDPK1 kinase activity by stimulating its dissociation from the 14-3-3 protein YWHAQ which acts as a negative regulator. May act as a tumor suppressor in colorectal carcinoma (PubMed:8752209).

Alternative names

MADH2, MADR2, SMAD2, Mothers against decapentaplegic homolog 2, MAD homolog 2, Mothers against DPP homolog 2, JV18-1, Mad-related protein 2, SMAD family member 2, hMAD-2, SMAD 2, Smad2, hSMAD2

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: EP784Y
Purification technique: Affinity purification Protein A
Specificity: This antibody is specific for MH 1 domain of Smad2.
Concentration

Storage

Shipped at conditions: Blue Ice
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

What is this antibody validated in?

Anti-Smad2 antibody [EP784Y] (ab40855) is a rabbit recombinant monoclonal antibody and is validated for use in Western Blot (WB), Flow Cytometry (Intra), Flow Cytometry (Flow Cyt), Immunoprecipitation (IP), Immunohistochemistry (IHC-P), Immunocytochemistry/immunofluorescence (ICC/IF) in Human, Rat samples.

What is the molecular weight of Smad2?

Anti-Smad2 [EP784Y] (ab40855) specifically detects a band for Smad2 (UniProt: Q15796) at a molecular weight of 58kDa.

Trusted by the scientific community

Anti-Smad2 [EP784Y] (ab40855) was first used in a scientific publication in 2006 and has been cited over 100 times in peer-reviewed journals.

Trial sizes available!

Test your antibody or perform pre-screening before committing to a larger quantity. Sold in 10µl. Discover our selection of trial-size antibodies.

Specificity confirmed

The specificity of Anti-Smad2 antibody [EP784Y] (ab40855) has been confirmed by Western blot testing in SMAD2 Knockout HeLa cells.

Other related products

We have a range of other formats of antibody clone [EP784Y] also available for your convenience:
ab40855, Carrier free - ab157371, PE - ab212096, Alexa Fluor® 488 - ab215913

Species reactivity
Mouse: We have preliminary internal testing data to indicate this antibody may not react with this species.
Please contact us for more information.

Patented technology
Our RabMAb^® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb^® patents.

What are the advantages of a recombinant monoclonal antibody?
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

Smad2 also known as Mothers against decapentaplegic homolog 2 (MAD2) or MADR2 is a signaling protein involved in the transforming growth factor-beta (TGF-Β) receptor pathway. Smad2 has a molecular mass of approximately 58 kDa and expresses in various tissues including epithelial mesenchymal and endothelial cells. Smad2 undergoes phosphorylation on serine residues in response to TGF-Β signaling converting it into phosphorylated forms often referred to as p-Smad2 or phospho-Smad2. These phosphorylated forms are critical for the relay of signals from the cell surface to the nucleus.

Biological function summary

Smad2 acts as an intracellular mediator for TGF-Β signaling a pathway important for regulating cell proliferation differentiation and apoptosis. Smad2 typically functions as part of a heteromeric complex with Smad4 another key player in TGF-Β signaling. Upon activation phosphorylated Smad2 combines with Smad4 to form a complex that translocates into the nucleus. This complex then binds to specific DNA sequences to regulate the transcription of target genes involved in processes such as cell growth inhibition and extracellular matrix production.

Pathways

Smad2 is integral to the TGF-Β and activin receptor signaling pathways. These pathways are essential in controlling cell growth and immune responses. Smad2 interacts with other proteins such as Smad3 in addition to Smad4 to modulate gene expression effectively. The interaction between Smad2 and these proteins ensures precise control of cellular responses to external stimuli emphasizing its pivotal role in maintaining cellular homeostasis.

Associated diseases and disorders

Smad2 correlates with various pathological conditions including fibrosis and cancer. Aberrant Smad2 signaling can contribute to the development of these diseases as excessive TGF-Β signaling promotes fibrotic tissue deposition and tumor progression. Smad2 connects with other proteins like Smad3 in these pathological contexts both acting as mediators of abnormal cellular behaviors. Understanding the regulatory mechanisms of Smad2 can help develop therapeutic strategies against disorders linked to dysregulated TGF-Β signaling.

Product promise

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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:
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13 product images

Immunoprecipitation - Anti-Smad2 antibody [EP784Y] (ab40855)
ab40855 (purified) at 1/20 immunoprecipitating EGFR in HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate.
Lane 1 (input): HeLa whole cell lysate (10μg)
Lane 2 (+): ab40855 + HeLa whole cell lysate.
Lane 3 (-): Rabbit monoclonal IgG (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) instead of ab40855 in HeLa whole cell lysate.
For western blotting, VeriBlot for IP Detection Reagent (HRP) ab131366 VeriBlot for IP (HRP) was used for detection (1/1000).
Blocking/Diluting buffer and concentration: 5% NFDM/TBST.
All lanes: Immunoprecipitation - Anti-Smad2 antibody [EP784Y] (ab40855)
Predicted band size: 52 kDa
Western blot - Anti-Smad2 antibody [EP784Y] (ab40855)
Lanes 1 - 3: Merged signal (red and green). Green - ab40855 observed at 58 kDa. Red - loading control, Anti-GAPDH antibody [mAbcam 9484] - Loading Control ab9484, observed at 37 kDa.

ab40855 was shown to specifically react with Smad2 in wild-type HeLa cells as signal was lost in Smad2 knockout cells. Wild-type and SMAD2 knockout samples were subjected to SDS-PAGE. ab40855 and Anti-GAPDH antibody [mAbcam 9484] - Loading Control ab9484 (Mouse anti-GAPDH loading control) were incubated overnight at 4°C at 1/10000 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-Smad2 antibody [EP784Y] (ab40855) at 1/1000 dilution
Lane 1: Wild-type HeLa whole cell lysate at 20 µg
Lane 2: Smad2 knockout HeLa whole cell lysate at 20 µg
Lane 3: A549 whole cell lysate at 20 µg
Predicted band size: 52 kDa
Western blot - Anti-Smad2 antibody [EP784Y] (ab40855)
Lanes 1 - 4: Merged signal (red and green). Green - ab40855 observed at 58 kDa. Red - loading control, Anti-GAPDH antibody [6C5] - Loading Control ab8245 observed at 37 kDa.
ab40855 was shown to react with Smad2 in wild-type HeLa. Loss of signal was observed when knockout cell line Human SMAD2 knockout HeLa cell line ab255430 (knockout cell lysate Human SMAD2 knockout HeLa cell lysate ab263833) was used. Wild-type and Smad2 knockout samples were subjected to SDS-PAGE. ab40855 and Anti-GAPDH antibody [6C5] - Loading Control (Anti-GAPDH antibody [6C5] - Loading Control ab8245) were incubated overnight at 4°C at 1 in 2000 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-Smad2 antibody [EP784Y] (ab40855) at 1/2000 dilution
Lane 1: A549 cell lysate at 20 µg
Lane 2: Jurkat cell lysate at 20 µg
Lane 2: Western blot - Human SMAD2 knockout HeLa cell line (Human SMAD2 knockout HeLa cell line ab255430)
Lane 3: Wild-type HeLa cell lysate at 20 µg
Lane 4: SMAD2 knockout HeLa cell lysate at 20 µg
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
Predicted band size: 52 kDa
Observed band size: 37 kDa, 55 kDa
Western blot - Anti-Smad2 antibody [EP784Y] (ab40855)
Diluting and blocking buffer: 5% NFDM /TBST
All lanes: Western blot - Anti-Smad2 antibody [EP784Y] (ab40855) at 1/2000 dilution
Lane 1: A-673 (Human muscle Ewing's Sarcoma cell line) whole cell lysate at 20 µg
Lane 2: HUVEC (Human umbilical vein endothelial cell line) whole cell lysate at 20 µg
Lane 3: C6 (Rat glial tumor cell line) whole cell 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: 52 kDa
Flow Cytometry (Intracellular) - Anti-Smad2 antibody [EP784Y] (ab40855)
ab40855 staining Smad2in the human cell line HeLa (Human epithelial cell line from cervix adenocarcinoma) by intracellular flow cytometry. Cells were fixed with 4% paraformaldehyde and the sample was incubated with the primary antibody at a dilution of 1/20. A goat anti rabbit IgG (Alexa Fluor^® 488) at a dilution of 1/2000 was used as the secondary antibody.
Isoytype control: Rabbit monoclonal IgG (Black)
Unlabelled control: Cell without incubation with primary antibody and secondary antibody (Blue)
Western blot - Anti-Smad2 antibody [EP784Y] (ab40855)
Blocking and diluting buffer: 5% NFDM/TBST
All lanes: Western blot - Anti-Smad2 antibody [EP784Y] (ab40855) at 1/10000 dilution
Lane 1: Jurkat (Human T cell leukemia cell line from peripheral blood) whole cell lysate at 20 µg
Lanes 2 - 3: HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell 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 20000 µg
Predicted band size: 52 kDa
Immunocytochemistry/ Immunofluorescence - Anti-Smad2 antibody [EP784Y] (ab40855)
ab40855 staining Smad2 in HeLa (human cervix adenocarcinoma) cells by ICC/IF (Immunocytochemistry/immunofluorescence). Cells were fixed with 4% Paraformaldehyde and permeabilized with 0.1% Triton X-100. Samples were incubated with primary antibody at a dilution of 1/100. A goat anti rabbit IgG (Alexa Fluor® 488) (Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077) was used as the secondary antibody at a dilution of 1/1000. Alexa Fluor® 594 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker ab195889 was used as a counterstain for primary antibody ab40855 at 1/1000. DAPI was used as a nuclear counterstain and PBS as a negative control.
Immunocytochemistry/ Immunofluorescence - Anti-Smad2 antibody [EP784Y] (ab40855)
Immunofluorescence staining of HeLa cells with purified ab40855 at a working dilution of 1/500, counter-stained with DAPI. The secondary antibody was an Alexa Fluor^®488 conjugated goat anti-rabbit (Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077), used at a dilution of 1/1000. The cells were fixed in 4% PFA and permeabilized using 0.1% Triton X 100. The negative control is shown in bottom right hand panel - for the negative control, PBS was used instead of the primary antibody.
Flow Cytometry (Intracellular) - Anti-Smad2 antibody [EP784Y] (ab40855)
Overlay histogram showing PC3 cells stained with ab40855 (red line). The cells were fixed with 80% methanol (5 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab40855, 1/100 dilution) for 30 min at 22°C. The secondary antibody used was Alexa Fluor^® 488 goat anti-rabbit IgG (H&L) (Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077) at 1/2000 dilution for 30 min at 22°C. Isotype control antibody (black line) was rabbit IgG (monoclonal) (1μg/1x10⁶ cells) used under the same conditions. Unlabelled sample (blue line) was also used as a control. Acquisition of >5,000 events were collected using a 20mW Argon ion laser (488nm) and 525/30 bandpass filter.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Smad2 antibody [EP784Y] (ab40855)
ab40855 stainingSmad2 in human bladder carcinoma tissue sections by Immunohistochemistry (IHC-P - paraformaldehyde-fixed, paraffin-embedded sections). Tissue was fixed with paraformaldehyde and antigen retrieval was by heat mediation in a EDTA buffer. Samples were incubated with primary antibody at a dilution of 1/50. A ImmunoHistoProbe one step HRP Polymer was used as a secondary antibody, ready to use.
Negative control 1: PBS in place of primary antibody.
Western blot - Anti-Smad2 antibody [EP784Y] (ab40855)
All lanes: Western blot - Anti-Smad2 antibody [EP784Y] (ab40855) at 1/500000 dilution
All lanes: Jurkat cell lysate
Predicted band size: 52 kDa
Observed band size: 58 kDa
Western blot - Anti-Smad2 antibody [EP784Y] (ab40855)
The molecular weight observed is consistent with what has been described in the literature (PMID: 34655614).
The up-regulation of pSmad2 is induced by TGF-beta treatment (PMID: 34655614, 24347165).
In Western blot, Anti-Smad2 (phospho S465 + S467) antibody [EPR28610-44] ab316117 was shown to bind specifically to Smad2. Target of interest was observed at 60 kDa in wild-type Hela cell lysates (lanes 2) with no signal observed at this size in Smad2 knockout cell line lysates (lanes 3-4 KO cell line Human SMAD2 knockout HeLa cell line ab255430/KO cell lysate Human SMAD2 knockout HeLa cell lysate ab263833 ).
In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (Anti-GAPDH antibody [EPR16891] - Loading Control ab181602) staining at 1/200000 dilution.
In Western blot, Anti-Smad2 antibody [EP784Y] - Total protein control (ab40855) staining at 1/1000 dilution.
All lanes: Western blot - Anti-Smad2 (phospho S465 + S467) antibody [EPR28610-44] (Anti-Smad2 (phospho S465 + S467) antibody [EPR28610-44] ab316117) at 1/1000 dilution
Lane 1: Untreated wild-type HeLa (human cervical adenocarcinoma epithelial cell) whole cell lysate (untreated membrane) at 20 µg with 5% NFDM/TBST
Lane 2: Wild-type Hela treated with 20 ng/ml TGF beta 1 for 60 minutes, whole cell lysate (untreated membrane) at 20 µg with 5% NFDM/TBST
Lane 3: Untreated Smad2 knockout Hela whole cell lysate (untreated membrane) at 20 µg with 5% NFDM/TBST
Lane 4: Smad2 knockout Hela treated with 20 ng/ml TGF beta 1 for 60 minutes, whole cell lysate (untreated membrane) at 20 µg with 5% NFDM/TBST
Lane 5: Untreated wild-type HeLa (human cervical adenocarcinoma epithelial cell) whole cell lysate (alkaline phosphatase treated membrane) at 20 µg with 5% NFDM/TBST
Lane 6: Wild-type Hela treated with 20 ng/ml TGF beta 1 for 60 minutes, whole cell lysate (alkaline phosphatase treated membrane) at 20 µg with 5% NFDM/TBST
Lane 7: Untreated Smad2 knockout Hela whole cell lysate (alkaline phosphatase treated membrane) at 20 µg with 5% NFDM/TBST
Lane 8: Smad2 knockout Hela treated with 20 ng/ml TGF beta 1 for 60 minutes, whole cell lysate (alkaline phosphatase treated membrane) at 20 µg with 5% NFDM/TBST
Secondary
All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/100000 dilution
Performed under reducing conditions.
Observed band size: 60 kDa, 36 kDa
Exposure time: 37s
ChIC/CUT&RUN sequencing - Anti-Smad2 antibody [EP784Y] (ab40855)
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 ab40855 [EP784Y]. 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.