For the best experience on the Abcam website please upgrade to a modern browser such as Google Chrome

Hello. We're improving abcam.com and we'd welcome your feedback.

Hello. We're improving abcam.com and we'd welcome your feedback.

Infomation icon

We haven't added this to the BETA yet

New BETA website

New BETA website

Hello. We're improving abcam.com and we'd welcome your feedback.

Take a look at our BETA site and see what we’ve done so far.

Switch on our new BETA site

Now available

Search and browse selected products

  • A selection of primary antibodies

Purchase these through your usual distributor

In the coming months

  • Additional product types
  • Supporting content
  • Sign in to your account
  • Purchase online
United States
Your country/region is currently set to:

If incorrect, please enter your country/region into the box below, to view site information related to your country/region.

Call (888) 77-ABCAM (22226) or contact us
Need help? Contact us

  • My account
  • Sign out
Sign in or Register with us

Welcome

Sign in or

Don't have an account?

Register with us
My basket
Quick order
Abcam homepage

  • Research Products
    By product type
    Primary antibodies
    Secondary antibodies
    ELISA and Matched Antibody Pair Kits
    Cell and tissue imaging tools
    Cellular and biochemical assays
    Proteins and Peptides
    By product type
    Proteomics tools
    Agonists, activators, antagonists and inhibitors
    Cell lines and Lysates
    Multiplex miRNA assays
    Multiplex Assays
    By research area
    Cancer
    Cardiovascular
    Cell Biology
    Epigenetics
    Metabolism
    Developmental Biology
    By research area
    Immunology
    Microbiology
    Neuroscience
    Signal Transduction
    Stem Cells
  • Customized Products & Partnerships
    Customized Products & Partnerships

    Customized products and commercial partnerships to accelerate your diagnostic and therapeutic programs.

    Customized products

    Partner with us

  • Support
    Support hub

    Access advice and support for any research roadblock

    View support hub

    Protocols

    Your experiments laid out step by step

    View protocols

  • Events
    • Conference calendar
    • Cancer
    • Cardiovascular
    • Epigenetics & Nuclear signaling
    • Immunology
    • Neuroscience
    • Stem cells
    • Tradeshows
    • Scientific webinars
    Keep up to date with the latest events

    Full event breakdown with abstracts, speakers, registration and more

    View global event calendar

  • Pathways
    Cell signalling pathways

    View all pathways

    View all interactive pathways

Explore the power of knock-out cell lines for your research

  1. Link

    smad3-ps423s425-elisa-kit-ab186038.pdf

  1. Send me a copy of this email
    I agree to the terms and conditions.
Signal Transduction Signaling Pathway Nuclear Signaling SMADs
Share by email
SimpleStep

SMAD3 (pS423/S425) ELISA Kit (ab186038)

  • Datasheet
  • SDS
  • Protocol Booklet
Submit a review Submit a question References (4)

Product price, shipping and contact information

Currently unavailable

Sorry, we can't display this right now.

Please contact us to place your order, or try again later.

 

Loading size & price…

 

Shipping and order information

Shipping info

Promotion Information

Abpromise

Guaranteed product quality, expert customer support.

Find out more.

Other - SMAD3 (pS423/S425) ELISA Kit (ab186038)
  • SMAD3 (pS423/S425) control lysate dilution series in 1X Cell Extraction Buffer PTR.
  • SMAD3 (pS423/S425) control lysate dilution series in 1X Cell Extraction Buffer PTR.
  • Linearity of dilution
  • SMAD3 (pS423/S425) phosphorylation in response to TGF-beta treatment.
  • SMAD3 (pS423/S425) phosphorylation in response to SB431542 treatment.
  • Example of a typical SMAD3 (pS423/S425) control lysate dilution series in 1X Cell Extraction Buffer PTR.

Key features and details

  • One-wash 90 minute protocol
  • Sensitivity: 10 µg/ml
  • Range: 10 µg/ml - 500 µg/ml
  • Sample type: Cell Lysate, Tissue Homogenate
  • Detection method: Colorimetric
  • Assay type: Semi-quantitative
  • Reacts with: Mouse, Human

You may also be interested in

Peptide
Smad3 peptide (ab204884)
Kit
Product image
BCA protein assay kit for low concentrations (ab207002)
ELISA
Product image
Human SMAD3 ELISA Kit (ab264624)

View more associated products

Overview

  • Product name

    SMAD3 (pS423/S425) ELISA Kit
    See all Smad3 kits
  • Detection method

    Colorimetric
  • Precision

    Intra-assay
    Sample n Mean SD CV%
    A431 extract 6 2.4%
    Inter-assay
    Sample n Mean SD CV%
    A431 3 7.4%
  • Sample type

    Cell Lysate, Tissue Homogenate
  • Assay type

    Semi-quantitative
  • Sensitivity

    10 µg/ml
  • Range

    10 µg/ml - 500 µg/ml
  • Assay time

    1h 30m
  • Assay duration

    One step assay
  • Species reactivity

    Reacts with: Mouse, Human
    Predicted to work with: Rat
  • Product overview

    Abcam’s SMAD3 (pS423/S425) in vitro SimpleStep ELISA™ (Enzyme-Linked Immunosorbent Assay) kit is designed for the semi-quantitative measurement of SMAD3 (pS423/S425) protein in human and mouse cells.


    The SimpleStep ELISA™ employs a labeled capture and detector antibody which immunocaptures the sample analyte in solution. This entire complex (capture antibody/protein/detector antibody) is in turn immobilized in the well by immunoaffinity via the anti-tag antibody. Samples or standards are added to the wells, followed by the antibody mix. After incubation, the wells are washed to remove unbound material; the TMB substrate is then added. The reaction is stopped by addition of Stop Solution which stops the color development and completes any color change from blue to yellow. Signal is generated proportionally to the amount of bound analyte and the intensity is measured at 450 nm. Optionally, instead of the endpoint reading, development of TMB can be recorded kinetically at 600 nm.


    ASSAY SPECIFICITY


    The SMAD3 (pS423/S425) assay detects endogenous levels of SMAD3 (GenBank Accession NP_001138574) in cellular lysates, only when phosphorylated at Ser423/425. Based on sequence similarity, cross reaction to SMAD2 may occur.


    SPECIES REACTIVITY


    This kit detects SMAD3 (pS423/S425) in Human and mouse cell culture extracts. Detection in rat samples is also expected. Other species should be tested on a case-by-case basis.


    Serum and plasma samples have not been tested with this kit.

  • Notes

    Abcam has not and does not intend to apply for the REACH Authorisation of customers’ uses of products that contain European Authorisation list (Annex XIV) substances.
    It is the responsibility of our customers to check the necessity of application of REACH Authorisation, and any other relevant authorisations, for their intended uses.

  • Platform

    Microplate

Properties

  • Storage instructions

    Store at +4°C. Please refer to protocols.
  • Components 1 x 96 tests
    10X Wash Buffer PT 1 x 15ml
    50X Cell Extraction Enhancer Solution 1 x 1ml
    5X Cell Extraction Buffer PTR 1 x 12ml
    Lyophilized SMAD3 (pS423/S425) Control Lysate 1 vial
    Plate Seal 1 unit
    SimpleStep Pre-Coated 96-Well Microplate (ab206978) 1 unit
    SMAD3 (pS423/S425) Capture Antibody 1 x 3ml
    SMAD3 (pS423/S425) Detector Antibody 1 x 3ml
    Stop Solution 1 x 12ml
    TMB Substrate 1 x 12ml
  • Research areas

    • Signal Transduction
    • Signaling Pathway
    • Nuclear Signaling
    • SMADs
    • Epigenetics and Nuclear Signaling
    • Nuclear Signaling Pathways
    • SMADs
    • Stem Cells
    • Signaling Pathways
    • TGF beta
    • Cytoplasmic
    • Epigenetics and Nuclear Signaling
    • Transcription
    • Transcription Factors
    • Cancer
    • Signal transduction
    • Nuclear signaling
    • SMAD family
    • Cancer
    • Cancer Metabolism
    • Response to hypoxia
    • Kits/ Lysates/ Other
    • Kits
    • ELISA Kits
    • ELISA Kits
    • Phosphoprotein ELISA kits
    • Metabolism
    • Pathways and Processes
    • Metabolism processes
    • Hypoxia
  • 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.
  • Involvement in disease

    Colorectal cancer
    Loeys-Dietz syndrome 3
  • Sequence similarities

    Belongs to the dwarfin/SMAD family.
    Contains 1 MH1 (MAD homology 1) domain.
    Contains 1 MH2 (MAD homology 2) domain.
  • Domain

    The MH1 domain is required for DNA binding. Also binds zinc ions which are necessary for the DNA binding.
    The MH2 domain is required for both homomeric and heteromeric interactions and for transcriptional regulation. Sufficient for nuclear import.
    The linker region is required for the TGFbeta-mediated transcriptional activity and acts synergistically with the MH2 domain.
  • Post-translational
    modifications

    Phosphorylated on serine and threonine residues. Enhanced phosphorylation in the linker region on Thr-179, Ser-204 and Ser-208 on EGF and TGF-beta treatment. Ser-208 is the main site of MAPK-mediated phosphorylation. CDK-mediated phosphorylation occurs in a cell-cycle dependent manner and inhibits both the transcriptional activity and antiproliferative functions of SMAD3. This phosphorylation is inhibited by flavopiridol. Maximum phosphorylation at the G(1)/S junction. Also phosphorylated on serine residues in the C-terminal SXS motif by TGFBR1 and ACVR1. TGFBR1-mediated phosphorylation at these C-terminal sites is required for interaction with SMAD4, nuclear location and transactivational activity, and appears to be a prerequisite for the TGF-beta mediated phosphorylation in the linker region. Dephosphorylated in the C-terminal SXS motif by PPM1A. This dephosphorylation disrupts the interaction with SMAD4, promotes nuclear export and terminates TGF-beta-mediated signaling. Phosphorylation at Ser-418 by CSNK1G2/CK1 promotes ligand-dependent ubiquitination and subsequent proteasome degradation, thus inhibiting SMAD3-mediated TGF-beta responses. Phosphorylated by PDPK1.
    Acetylation in the nucleus by EP300 in the MH2 domain regulates positively its transcriptional activity and is enhanced by TGF-beta.
    Ubiquitinated. Monoubiquitinated, leading to prevent DNA-binding. Deubiquitination by USP15 alleviates inhibition and promotes activation of TGF-beta target genes.
    Poly-ADP-ribosylated by PARP1 and PARP2. ADP-ribosylation negatively regulates SMAD3 transcriptional responses during the course of TGF-beta signaling.
  • 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 (PubMed:15799969). Through the action of the phosphatase PPM1A, released from the SMAD2/SMAD4 complex, and exported out of the nucleus by interaction with RANBP1 (PubMed:16751101, PubMed:19289081). Co-localizes with LEMD3 at the nucleus inner membrane (PubMed:15601644). MAPK-mediated phosphorylation appears to have no effect on nuclear import (PubMed:19218245). PDPK1 prevents its nuclear translocation in response to TGF-beta (PubMed:17327236).
  • Target information above from: UniProt accession P84022 The UniProt Consortium
    The Universal Protein Resource (UniProt) in 2010
    Nucleic Acids Res. 38:D142-D148 (2010) .

    Information by UniProt
  • Alternative names

    • DKFZP586N0721
    • DKFZp686J10186
    • hMAD 3
    • hMAD-3
    • hSMAD3
    • HSPC193
    • HST17436
    • JV15 2
    • JV15-2
    • JV152
    • LDS1C
    • LDS3
    • MAD (mothers against decapentaplegic Drosophila) homolog 3
    • MAD homolog 3
    • Mad homolog JV15 2
    • Mad protein homolog
    • MAD, mothers against decapentaplegic homolog 3
    • Mad3
    • MADH 3
    • MADH3
    • MGC60396
    • Mothers against decapentaplegic homolog 3
    • Mothers against DPP homolog 3
    • SMA and MAD related protein 3
    • SMAD
    • SMAD 3
    • SMAD family member 3
    • SMAD, mothers against DPP homolog 3
    • Smad3
    • SMAD3_HUMAN
    see all
  • Database links

    • Entrez Gene: 4088 Human
    • Entrez Gene: 17127 Mouse
    • Entrez Gene: 25631 Rat
    • Omim: 603109 Human
    • SwissProt: P84022 Human
    • SwissProt: Q8BUN5 Mouse
    • SwissProt: P84025 Rat
    • Unigene: 727986 Human
    • Unigene: 742270 Human
    • Unigene: 7320 Mouse
    • Unigene: 10636 Rat
    see all

Associated products

  • Related Products

    • RepSox, TGF-beta type 1 receptor inhibitor (ab142139)

Images

  • Other - SMAD3 (pS423/S425) ELISA Kit (ab186038)
    Other - SMAD3 (pS423/S425) ELISA Kit (ab186038)

    SimpleStep ELISA technology allows the formation of the antibody-antigen complex in one single step, reducing assay time to 90 minutes. Add samples or standards and antibody mix to wells all at once, incubate, wash, and add your final substrate. See protocol for a detailed step-by-step guide.

     

  • SMAD3 (pS423/S425) control lysate dilution series in 1X Cell Extraction Buffer PTR.
    SMAD3 (pS423/S425) control lysate dilution series in 1X Cell Extraction Buffer PTR.

    Raw data values are shown in the table. 

  • SMAD3 (pS423/S425) control lysate dilution series in 1X Cell Extraction Buffer PTR.
    SMAD3 (pS423/S425) control lysate dilution series in 1X Cell Extraction Buffer PTR.

    Example of a typical SMAD3 (pS423/S425) control lysate dilution series in 1X Cell Extraction Buffer PTR.  The SMAD3 lysate dilution series was prepared as described.  Raw data values are shown in the table. 

  • Linearity of dilution
    Linearity of dilution

    Linearity of dilution in representative sample matrices. Cellular lysates were prepared at 3 concentrations in common media containing 1X Cell Extraction Buffer PTR. Data from duplicate measurements of SMAD3 (pS423/S425) are normalized and plotted.

  • SMAD3 (pS423/S425) phosphorylation in response to TGF-beta treatment.
    SMAD3 (pS423/S425) phosphorylation in response to TGF-beta treatment.

    Induction of SMAD3 (pS423/S425) phosphorylation in HeLa cells in response to TGF-β treatment. HeLa cells were cultured in 96-well tissue culture plates, serum-starved and treated (60 min) with a dose-range of TGF-β before cell lysis. Data from quadruplicate measurements of SMAD3 (pS423/S425) are plotted.

  • SMAD3 (pS423/S425) phosphorylation in response to SB431542 treatment.
    SMAD3 (pS423/S425) phosphorylation in response to SB431542 treatment.

    Inhibition of SMAD3 (pS423/S425) phosphorylation in HeLa cells in response to SB431542 treatment. HeLa cells were cultured in 96-well tissue culture plates and treated with a dose-range of SB431542 (60 min). Cells were then stimulated with TGF-β (60 min) and lysed. Data from quadruplicate measurements of SMAD3 (pS423/S425) are plotted.

  • Example of a typical SMAD3 (pS423/S425) control lysate dilution series in 1X Cell Extraction Buffer PTR.
    Example of a typical SMAD3 (pS423/S425) control lysate dilution series in 1X Cell Extraction Buffer PTR.

    The SMAD3 lysate dilution series was prepared as described.  Raw data values are shown in the table. 

    Kit Control lysates are provided at a concentration that give consistent signal between different lots. Lysates are produced and formulated by signal intensity to be consistent to within 30% of the previous lot. As such, Control lysates are not provided with a protein concentration.  

Protocols

  • Protocol Booklet

Click here to view the general protocols

Datasheets and documents

  • SDS download

  • Datasheet download

    Download

References (4)

Publishing research using ab186038? Please let us know so that we can cite the reference in this datasheet.

ab186038 has been referenced in 4 publications.

  • Guan R  et al. Chitinase-like protein YKL-40 regulates human bronchial epithelial cells proliferation, apoptosis, and migration through TGF-ß1/Smads pathway. Hum Exp Toxicol 39:451-463 (2020). PubMed: 31797699
  • Hasan AA  et al. Mechanisms of GLP-1 receptor-independent renoprotective effects of the dipeptidyl peptidase type 4 inhibitor linagliptin in GLP-1 receptor knockout mice with 5/6 nephrectomy. Kidney Int 95:1373-1388 (2019). PubMed: 30979564
  • Tanino Y  et al. Syndecan-4 Inhibits the Development of Pulmonary Fibrosis by Attenuating TGF-ß Signaling. Int J Mol Sci 20:N/A (2019). PubMed: 31600983
  • Kudipudi PK  et al. Betaglycan (TßRIII) is a Key Factor in TGF-ß2 Signaling in Prepubertal Rat Sertoli Cells. Int J Mol Sci 20:N/A (2019). PubMed: 31835434

Customer reviews and Q&As

Show All Reviews Q&A
Submit a review Submit a question

There are currently no Customer reviews or Questions for ab186038.
Please use the links above to contact us or submit feedback about this product.

Please note: All products are "FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC PROCEDURES"
For licensing inquiries, please contact partnerships@abcam.com

Get resources and offers direct to your inbox Sign up
A-Z by research area
  • Cancer
  • Cardiovascular
  • Cell biology
  • Developmental biology
  • Epigenetics & Nuclear signaling
  • Immunology
  • Metabolism
  • Microbiology
  • Neuroscience
  • Signal transduction
  • Stem cells
A-Z by product type
  • Primary antibodies
  • Secondary antibodies
  • Biochemicals
  • Isotype controls
  • Flow cytometry multi-color selector
  • Kits
  • Loading controls
  • Lysates
  • Peptides
  • Proteins
  • Slides
  • Tags and cell markers
  • Tools & Reagents
Help & support
  • Support
  • Make an Inquiry
  • Protocols & troubleshooting
  • Placing an order
  • RabMAb products
  • Biochemical product FAQs
  • Training
  • Browse by Target
Company
  • Corporate site
  • Investor relations
  • Company news
  • Careers
  • About us
  • Blog
Events
  • Tradeshows
  • Conferences
International websites
  • abcam.cn
  • abcam.co.jp

Join with us

  • LinkedIn
  • facebook
  • Twitter
  • YouTube
  • Terms of sale
  • Website terms of use
  • Cookie policy
  • Privacy policy
  • Legal
  • Modern slavery statement
© 1998-2022 Abcam plc. All rights reserved.