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    smad2-phospho-s255-antibody-epr2856n-ab188334.pdf

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Signal Transduction Signaling Pathway Nuclear Signaling SMADs
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RecombinantRabMAb

Recombinant Anti-Smad2 (phospho S255) antibody [EPR2856(N)] (ab188334)

  • Datasheet
  • Certificate of Compliance
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Western blot - Anti-Smad2 (phospho S255) antibody [EPR2856(N)] (ab188334)
  • Western blot - Anti-Smad2 (phospho S255) antibody [EPR2856(N)] (ab188334)
  • Western blot - Anti-Smad2 (phospho S255) antibody [EPR2856(N)] (ab188334)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Smad2 (phospho S255) antibody [EPR2856(N)] (ab188334)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Smad2 (phospho S255) antibody [EPR2856(N)] (ab188334)
  • Dot Blot - Anti-Smad2 (phospho S255) antibody [EPR2856(N)] (ab188334)
  • Immunoprecipitation - Anti-Smad2 (phospho S255) antibody [EPR2856(N)] (ab188334)
  • 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

Conjugates logo Related conjugates and formulations

Carrier Free

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Recombinant Human Smad2 protein (ab63176)

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Overview

  • 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.

  • 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.
  • 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
    For more information see here.

    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

  • 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

    • Signal Transduction
    • Signaling Pathway
    • Nuclear Signaling
    • SMADs
    • Epigenetics and Nuclear Signaling
    • Nuclear Signaling Pathways
    • SMADs
    • Stem Cells
    • Signaling Pathways
    • TGF beta
    • Cytoplasmic
    • Cancer
    • Growth factors
    • TGF

Associated products

  • Alternative Versions

    • Anti-Smad2 (phospho S255) antibody [EPR2856(N)] - BSA and Azide free (ab219598)
  • Compatible Secondaries

    • Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077)
    • Goat Anti-Rabbit IgG H&L (HRP) (ab205718)
  • Isotype control

    • Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730)
  • Positive Controls

    • HeLa Ser/Thr Phosphatase-Inhibited Whole Cell Lysate Set: Calyculin-Treated and Vehicle-Treated (ab135395)
  • Recombinant Protein

    • Recombinant Human Smad2 protein (ab63176)

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.
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.
IP
1/50 - 1/70.
WB
1/1000 - 1/10000. Detects a band of approximately 58 kDa (predicted molecular weight: 52 kDa).
Notes
Dot blot
Use at an assay dependent concentration.
IHC-P
1/50 - 1/100. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
IP
1/50 - 1/70.
WB
1/1000 - 1/10000. Detects a band of approximately 58 kDa (predicted molecular weight: 52 kDa).

Target

  • 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
    modifications

    Phosphorylated 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.
  • Target information above from: UniProt accession Q15796 The UniProt Consortium
    The Universal Protein Resource (UniProt) in 2010
    Nucleic Acids Res. 38:D142-D148 (2010) .

    Information by UniProt
  • 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
    • Unigene: 705764 Human
    • Unigene: 391091 Mouse
    • Unigene: 2755 Rat
    see all
  • Alternative names

    • Drosophila, homolog of, MADR2 antibody
    • hMAD-2 antibody
    • HsMAD2 antibody
    • JV18 antibody
    • JV18-1 antibody
    • JV181 antibody
    • MAD antibody
    • MAD homolog 2 antibody
    • MAD Related Protein 2 antibody
    • Mad-related protein 2 antibody
    • MADH2 antibody
    • MADR2 antibody
    • MGC22139 antibody
    • MGC34440 antibody
    • Mother against DPP homolog 2 antibody
    • Mothers against decapentaplegic homolog 2 antibody
    • Mothers against decapentaplegic, Drosophila, homolog of, 2 antibody
    • Mothers against DPP homolog 2 antibody
    • OTTHUMP00000163489 antibody
    • Sma and Mad related protein 2 antibody
    • Sma- and Mad-related protein 2 MAD antibody
    • SMAD 2 antibody
    • SMAD family member 2 antibody
    • SMAD, mothers against DPP homolog 2 antibody
    • SMAD2 antibody
    • SMAD2_HUMAN antibody
    see all

Images

  • Western blot - Anti-Smad2 (phospho S255) antibody [EPR2856(N)] (ab188334)
    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)
    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)
    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)
    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.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Smad2 (phospho S255) antibody [EPR2856(N)] (ab188334)
    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.

  • Dot Blot - Anti-Smad2 (phospho S255) antibody [EPR2856(N)] (ab188334)
    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 .

  • Immunoprecipitation - Anti-Smad2 (phospho S255) antibody [EPR2856(N)] (ab188334)
    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

  • Anti-Smad2 (phospho S255) antibody [EPR2856(N)] (ab188334)
    Anti-Smad2 (phospho S255) antibody [EPR2856(N)] (ab188334)

Protocols

  • Western blot protocols
  • Immunoprecipitation protocols
  • Immunohistochemistry protocols

Click here to view the general protocols

Datasheets and documents

  • Datasheet download

    Download

Certificate of Compliance

To download a Certificate of Compliance, please enter your Lot number below:

References (40)

Publishing research using ab188334? Please let us know so that we can cite the reference in this datasheet.

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
View all Publications for this product

Customer reviews and Q&As

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1-2 of 2 Abreviews or Q&A

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) abreview for Anti-Smad2 (phospho S255) antibody [EPR2856(N)]

Excellent
Abreviews
Abreviews
abreview image
Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Human Tissue sections (NSCLC: A549 cells)
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: pH6
Permeabilization
No
Specification
NSCLC: A549 cells
Blocking step
(agent) for 10 minute(s) · Concentration: 100% · Temperature: 22°C
Fixative
Paraformaldehyde
Read More

Abcam user community

Verified customer

Submitted Nov 24 2020

Immunocytochemistry/ Immunofluorescence abreview for Anti-Smad2 (phospho S255) antibody [EPR2856(N)]

Excellent
Abreviews
Abreviews
abreview image
Application
Immunocytochemistry/ Immunofluorescence
Sample
Hamster Cell (ovary)
Permeabilization
No
Specification
ovary
Blocking step
goat serum/0.2% triton X100 as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 10% · Temperature: 25°C
Fixative
Paraformaldehyde
Read More

Prof. Shyamal Roy

Verified customer

Submitted Jun 02 2016

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