Anti-Smad2 (phospho S467) antibody (ab53100)

Rabbit polyclonal Smad2 (phospho S467) antibody. Validated in WB, ELISA, IHC and tested in Mouse, Rat, Human. Cited in 32 publication(s). Independently reviewed in 14 review(s).

Overview

  • Product name

    Anti-Smad2 (phospho S467) antibody
    See all Smad2 primary antibodies
  • Description

    Rabbit polyclonal to Smad2 (phospho S467)
  • Host species

    Rabbit
  • Specificity

    Detects endogenous levels of Smad2 only when phosphorylated at serine 467
  • Tested applications

    Suitable for: WB, ELISA, IHC-Pmore details
  • Species reactivity

    Reacts with: Mouse, Rat, Human
  • Immunogen

    Synthetic peptide corresponding to Human Smad2.
    (Peptide available as ab138919)

  • Positive control

    • HeLa cells treated with PMA

Properties

Applications

Our Abpromise guarantee covers the use of ab53100 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
WB 1/300 - 1/1000. Detects a band of approximately 58 kDa (predicted molecular weight: 58 kDa).
ELISA 1/5000.
IHC-P 1/50 - 1/100. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

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.
  • Information by UniProt
  • Database links

  • 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

  • All lanes : Anti-Smad2 (phospho S467) antibody (ab53100) at 1/300 dilution

    Lane 1 : HeLa cell extract treated with PMA (125ng/ml, 15 mins)
    Lane 2 : HeLa cell extract treated with PMA (125ng/ml, 15 mins) with phosphopeptide

    Predicted band size: 58 kDa
    Observed band size: 58 kDa

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human breast carcinoma tissue labeling Smad2 (phospho S467) with ab53100. The image on the right is blocked with a phosphopeptide.

     

  • Anti-Smad2 (phospho S467) antibody (ab53100) at 1/750 dilution (for 16 hours at 4°C) + Mouse brain whole tissue lysate at 18 µg

    Secondary
    An HRP-conjugated Goat anti-rabbit-IgG polyclonal at 1/7000 dilution

    Developed using the ECL technique.

    Performed under reducing conditions.

    Predicted band size: 58 kDa
    Observed band size: 58 kDa


    Exposure time: 2 minutes


    Blocking Step: 5% Milk for 1 hour at 18°C

    See Abreview

  • Anti-Smad2 (phospho S467) antibody (ab53100) at 1/1500 dilution + Rat lung whole tissue lysate at 15 µg with Milk at 5 %

    Secondary
    Goat antirabbit conjugated to HRP at 1/7000 dilution

    Developed using the ECL technique.

    Performed under reducing conditions.

    Predicted band size: 58 kDa
    Additional bands at: 59 kDa. We are unsure as to the identity of these extra bands.


    Exposure time: 2 minutes

    See Abreview

  • ab53100 detecting phospho-Smad2 in Mouse brain samples by indirect ELISA. Samples were blocked with 5% BSA for 1 hour at 20°C and incubated with the primary antibody (1/750) for 4 hours at 20°C. An HRP-conjugated Goat anti-rabbit IgG polyclonal (1/5000) was used as the secondary antibody.

    See Abreview

References

This product has been referenced in:

  • Zhou B  et al. Long Non-Coding RNA TFAP2A-AS1 Inhibits Cell Proliferation and Invasion in Breast Cancer via miR-933/SMAD2. Med Sci Monit 25:1242-1253 (2019). Read more (PubMed: 30768589) »
  • Zhao H  et al. PDCD5 inhibits osteosarcoma cell metastasis via targeting TGF-ß1/Smad signaling pathway and is associated with good prognosis. Am J Transl Res 11:1116-1128 (2019). Read more (PubMed: 30899411) »
See all 41 Publications for this product

Customer reviews and Q&As

1-10 of 32 Abreviews or Q&A

Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Human Tissue sections (liver)
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: citric buffer
Permeabilization
Yes - Tx100
Specification
liver
Blocking step
Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 10% · Temperature: 25°C
Fixative
Paraformaldehyde

Abcam user community

Verified customer

Submitted Sep 30 2019

Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Mouse Tissue sections (liver)
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: citric buffer
Permeabilization
Yes - Tx100
Specification
liver
Blocking step
Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 10%
Fixative
Paraformaldehyde

Abcam user community

Verified customer

Submitted Sep 30 2019

Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Mouse Tissue sections (cartilage)
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: citric
Permeabilization
No
Specification
cartilage
Blocking step
Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 27°C
Fixative
Paraformaldehyde

Abcam user community

Verified customer

Submitted Jul 06 2019

Application
Immunohistochemistry (Frozen sections)
Sample
Mouse Tissue sections (embryo heart)
Permeabilization
Yes - 0.3% Triton
Specification
embryo heart
Blocking step
Normal Donkey Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 22°C
Fixative
Paraformaldehyde

Abcam user community

Verified customer

Submitted Mar 07 2018

Application
Western blot
Sample
Rat Tissue lysate - whole (Muscle)
Gel Running Conditions
Reduced Denaturing (9)
Loading amount
50 µg
Specification
Muscle
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 4°C

Mr. Baptiste Jude

Verified customer

Submitted Feb 26 2018

Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Mouse Tissue sections (Prostate)
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: Citrate, pH 6
Permeabilization
No
Specification
Prostate
Blocking step
Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 10% · Temperature: 20°C
Fixative
Paraformaldehyde

Abcam user community

Verified customer

Submitted Dec 29 2017

Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Human Tissue sections (Lung, colon cancer, normal colon, Glioblastoma)
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: Citrate, Tris EDTA
Permeabilization
No
Specification
Lung, colon cancer, normal colon, Glioblastoma
Blocking step
Milk as blocking agent for 1 hour(s) and 30 minute(s) · Concentration: 10% · Temperature: 25°C
Fixative
Formaldehyde

Sravya Palavalasa

Verified customer

Submitted May 16 2016

Application
Immunocytochemistry/ Immunofluorescence
Blocking step
Odyssey Blocking Buffer as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 100% · Temperature: 26°C
Sample
Human Cell (Ovarian Granulosa Cell Line)
Specification
Ovarian Granulosa Cell Line
Permeabilization
Yes - Ethanol and Triton
Fixative
Paraformaldehyde

Dr. Francesco Elia Marino

Verified customer

Submitted Jun 10 2014

Application
Western blot
Loading amount
40 µg
Gel Running Conditions
Reduced Non-Denaturing (Native) (12)
Sample
Mouse Cell lysate - whole cell (Gastrocnemius)
Specification
Gastrocnemius
Blocking step
Commercial Blocking Buffer as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 100% · Temperature: 26°C

Abcam user community

Verified customer

Submitted May 23 2014

Application
Western blot
Loading amount
60 µg
Gel Running Conditions
Reduced Non-Denaturing (Native) (12)
Sample
Human Cell lysate - nuclear (COV434 cell line)
Specification
COV434 cell line
Treatment
Activin A 10 ng/ml for 24 hours
Blocking step
Odyssey Blocking Buffer as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 100% · Temperature: 26°C

Dr. Francesco Elia Marino

Verified customer

Submitted Mar 21 2014

1-10 of 32 Abreviews or Q&A

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