Recombinant Anti-Smad2 (phospho S255) antibody [EPR2856(N)] (ab188334)
![Western blot - Anti-Smad2 (phospho S255) antibody [EPR2856(N)] (ab188334)](https://www.abcam.com/ps/products/188/ab188334/Images/ab188334-451258-AntiSmad2-phospho-S255-antibody-western-blot.jpg "Western blot - Anti-Smad2 (phospho S255) antibody [EPR2856(N)] (ab188334)")
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR2856(N)] to Smad2 (phospho S255)
- Suitable for: Dot blot, IHC-P, IP, WB
- Reacts with: Mouse, Rat, Human
Related conjugates and formulations
Overview
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Product name
Anti-Smad2 (phospho S255) antibody [EPR2856(N)]
See all Smad2 primary antibodies -
Description
Rabbit monoclonal [EPR2856(N)] to Smad2 (phospho S255) -
Host species
Rabbit -
Tested applications
Suitable for: Dot blot, IHC-P, IP, WBmore details -
Species reactivity
Reacts with: Mouse, Rat, Human -
Immunogen
Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.
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Positive control
- Hela treated with Okadaic acid and Calyculin A, Hela treated with Okadaic acid and Calyculin A, Human endometrium, Human transitional cell carcinoma of bladder. RAW 264.7 (Mouse Abelson murine leukemia virus-induced tumor macrophage) and PC-12 (Rat adrenal gland pheochromocytoma) whole cell lysates.
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General notes
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 production
Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.
Properties
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Form
Liquid -
Storage instructions
Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C long term. Avoid freeze / thaw cycle. -
Storage buffer
pH: 7.2
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol, 0.05% BSA -
Concentration information loading... -
Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
EPR2856(N) -
Isotype
IgG -
Research areas
Associated products
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Alternative Versions
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Compatible Secondaries
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Isotype control
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Positive Controls
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Recombinant Protein
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab188334 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
| Application | Abreviews | Notes |
|---|---|---|
| Dot blot |
Use at an assay dependent concentration.
|
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| IHC-P | (1) |
1/50 - 1/100. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
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| IP |
1/50 - 1/70.
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| WB |
1/1000 - 1/10000. Detects a band of approximately 58 kDa (predicted molecular weight: 52 kDa).
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| Notes |
|---|
|
Dot blot
Use at an assay dependent concentration. |
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IHC-P
1/50 - 1/100. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol. |
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IP
1/50 - 1/70. |
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WB
1/1000 - 1/10000. Detects a band of approximately 58 kDa (predicted molecular weight: 52 kDa). |
Target
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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. May act as a tumor suppressor in colorectal carcinoma. -
Tissue specificity
Expressed at high levels in skeletal muscle, heart and placenta. -
Sequence similarities
Belongs to the dwarfin/SMAD family.
Contains 1 MH1 (MAD homology 1) domain.
Contains 1 MH2 (MAD homology 2) domain. -
Post-translational
modificationsPhosphorylated on one or several of Thr-220, Ser-245, Ser-250, and Ser-255. In response to TGF-beta, phosphorylated on Ser-465/467 by TGF-beta and activin type 1 receptor kinases. Able to interact with SMURF2 when phosphorylated on Ser-465/467, recruiting other proteins, such as SNON, for degradation. In response to decorin, the naturally occurring inhibitor of TGF-beta signaling, phosphorylated on Ser-240 by CaMK2. Phosphorylated by MAPK3 upon EGF stimulation; which increases transcriptional activity and stability, and is blocked by calmodulin.
In response to TGF-beta, ubiquitinated by NEDD4L; which promotes its degradation.
Acetylated on Lys-19 by coactivators in response to TGF-beta signaling, which increases transcriptional activity. Isoform short: Acetylation increases DNA binding activity in vitro and enhances its association with target promoters in vivo. Acetylation in the nucleus by EP300 is enhanced by TGF-beta. -
Cellular localization
Cytoplasm. Nucleus. Cytoplasmic and nuclear in the absence of TGF-beta. On TGF-beta stimulation, migrates to the nucleus when complexed with SMAD4. On dephosphorylation by phosphatase PPM1A, released from the SMAD2/SMAD4 complex, and exported out of the nucleus by interaction with RANBP1. - Information by UniProt
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Database links
- Entrez Gene: 4087 Human
- Entrez Gene: 17126 Mouse
- Entrez Gene: 29357 Rat
- Omim: 601366 Human
- SwissProt: Q15796 Human
- SwissProt: Q62432 Mouse
- SwissProt: O70436 Rat
- Unigene: 12253 Human
see all -
Alternative names
- Drosophila, homolog of, MADR2 antibody
- hMAD-2 antibody
- HsMAD2 antibody
see all
Images
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Western blot - Anti-Smad2 (phospho S255) antibody [EPR2856(N)] (ab188334)All lanes : Anti-Smad2 (phospho S255) antibody [EPR2856(N)] (ab188334) at 1/10000 dilution
Lane 1 : PC-12 (Rat adrenal gland pheochromocytoma) whole cell lysate
Lane 2 : PC-12 (Rat adrenal gland pheochromocytoma) whole cell lysate, Then the membrane was incubated with phosphatase
Lysates/proteins at 15 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution (Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated)
Predicted band size: 52 kDa -
![Western blot - Anti-Smad2 (phospho S255) antibody [EPR2856(N)] (ab188334)](https://www.abcam.com/ps/products/188/ab188334/Images/ab188334-451257-AntiSmad2-phospho-S255-antibody-western-blot.jpg)
Western blot - Anti-Smad2 (phospho S255) antibody [EPR2856(N)] (ab188334)All lanes : Anti-Smad2 (phospho S255) antibody [EPR2856(N)] (ab188334) at 1/10000 dilution
Lane 1 : RAW 264.7 (Mouse Abelson murine leukemia virus-induced tumor macrophage) whole cell lysate
Lane 2 : RAW 264.7 (Mouse Abelson murine leukemia virus-induced tumor macrophage) whole cell lysate. Then the membrane was incubated with phosphatase.
Lysates/proteins at 15 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution (Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated)
Predicted band size: 52 kDa -
![Western blot - Anti-Smad2 (phospho S255) antibody [EPR2856(N)] (ab188334)](https://www.abcam.com/ps/products/188/ab188334/Images/ab188334-221861-anti-smad2-phospho-s255-antibody-epr2856n-western-blot.jpg)
Western blot - Anti-Smad2 (phospho S255) antibody [EPR2856(N)] (ab188334)All lanes : Anti-Smad2 (phospho S255) antibody [EPR2856(N)] (ab188334) at 1/10000 dilution
Lane 1 : Hela treated with Okadaic acid and Calyculin A
Lane 2 : Hela treated with Okadaic acid and Calyculin A , then treated with Lambda phosphorylase
Lane 3 : Untreated Hela
Lysates/proteins at 10 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated at 1/1000 dilution
Predicted band size: 52 kDa -
![Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Smad2 (phospho S255) antibody [EPR2856(N)] (ab188334)](https://www.abcam.com/ps/products/188/ab188334/Images/ab188334-221860-anti-smad2-phospho-s255-antibody-epr2856n-immunohistochemistry.jpg)
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Smad2 (phospho S255) antibody [EPR2856(N)] (ab188334)Immunohistochemical analysis of formalin fixed paraffin embedded Human endometrium labeling Smad2 (phospho S255) with ab188334 at 1/100 dilution and HRP Polymer for Rabbit IgG. Counterstained with Hematoxylin.
Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
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![Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Smad2 (phospho S255) antibody [EPR2856(N)] (ab188334)](https://www.abcam.com/ps/products/188/ab188334/Images/ab188334-221858-anti-smad2-phospho-s255-antibody-epr2856n-immunohistochemistry.jpg)
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Smad2 (phospho S255) antibody [EPR2856(N)] (ab188334)Immunohistochemical analysis of formalin fixed paraffin embedded Human transitional cell carcinoma of bladder labeling Smad2 (phospho S255) with ab188334 at 1/100 dilution and HRP Polymer for Rabbit IgG. Counterstained with Hematoxylin.
Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
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![Dot Blot - Anti-Smad2 (phospho S255) antibody [EPR2856(N)] (ab188334)](https://www.abcam.com/ps/products/188/ab188334/Images/ab188334-312021-anti-smad2-phospho-s255-antibody-epr2856n-dot-blot.jpg)
Dot Blot - Anti-Smad2 (phospho S255) antibody [EPR2856(N)] (ab188334)Dot blot analysis of Smad2 (S255) phospho peptide (Lane 1), Smad2 non-phospho peptide (Lane 2), labelling Smad2 (S255) phospho peptide with ab188334 at a dilution of 1:1000 dilution (1.365ug/ml). A Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (ab97051) was used as the secondary antibody at a dilution of 1:20,000 dilution. Blocking buffer: 5% NFDM/TBST. Dilution buffer: 5% NFDM /TBST .
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![Immunoprecipitation - Anti-Smad2 (phospho S255) antibody [EPR2856(N)] (ab188334)](https://www.abcam.com/ps/products/188/ab188334/Images/ab188334-221856-anti-smad2-phospho-s255-antibody-epr2856n-immunoprecipitation.jpg)
Immunoprecipitation - Anti-Smad2 (phospho S255) antibody [EPR2856(N)] (ab188334)Immunoprecipitation of Hela cells treated with Okadaic acis and Calyculin A (Lane 1) or PBS (Lane 2) labeling Smad2 (phospho S255) with ab188334 at 1/50 dilution and Anti-Rabbit IgG (HRP), specific to the non-reduced form of IgG at 1/1500 dilution
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![Anti-Smad2 (phospho S255) antibody [EPR2856(N)] (ab188334)](https://www.abcam.com/ps/products/188/ab188334/Images/ab188334-6-benefits-of-recombinant-antibodies.png)
Anti-Smad2 (phospho S255) antibody [EPR2856(N)] (ab188334)
Protocols
Datasheets and documents
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Datasheet download
Certificate of Compliance
References (40)
ab188334 has been referenced in 40 publications.
- Li Y et al. Palmitoylethanolamide (PEA) reduces postoperative adhesions after experimental strabismus surgery in rabbits by suppressing canonical and non-canonical TGFß signaling through PPARa. Biochem Pharmacol 184:114398 (2021). PubMed: 33385371
- Xie F et al. Adipose-derived mesenchymal stem cells inhibit cell proliferation and migration and suppress extracellular matrix synthesis in hypertrophic-scar and keloid fibroblasts. Exp Ther Med 21:139 (2021). PubMed: 33456506
- Wang W et al. Positive feedback regulation between USP15 and ERK2 inhibits osteoarthritis progression through TGF-ß/SMAD2 signaling. Arthritis Res Ther 23:84 (2021). PubMed: 33726807
- Wu SJ et al. Activation of the Cholinergic Anti-inflammatory Pathway Attenuated Angiotension II-Dependent Hypertension and Renal Injury. Front Pharmacol 12:593682 (2021). PubMed: 33815099
- Tian T et al. FAST1 Predicts Poor Survival of Renal Carcinoma and Promotes Its Progression Through the TGF-ß/Smad Pathway. Onco Targets Ther 14:1487-1499 (2021). PubMed: 33679133
