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

  1. Link

    products/primary-antibodies/smad2-phospho-s250-antibody-epr26263-58-ab300079.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
Validated using a knockout cell lineRecombinantRabMAb

Recombinant Anti-Smad2 (phospho S250) antibody [EPR26263-58] (ab300079)

  • Datasheet
  • SDS
  • Certificate of Compliance
Submit a review Submit a question

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.

Western blot - Anti-Smad2 (phospho S250) antibody [EPR26263-58] (AB300079)
  • Western blot - Anti-Smad2 (phospho S250) antibody [EPR26263-58] (AB300079)
  • Western blot - Anti-Smad2 (phospho S250) antibody [EPR26263-58] (AB300079)
  • Western blot - Anti-Smad2 (phospho S250) antibody [EPR26263-58] (AB300079)
  • Immunocytochemistry/ Immunofluorescence - Anti-Smad2 (phospho S250) antibody [EPR26263-58] (AB300079)
  • Immunoprecipitation - Anti-Smad2 (phospho S250) antibody [EPR26263-58] (AB300079)
  • Immunoprecipitation - Anti-Smad2 (phospho S250) antibody [EPR26263-58] (AB300079)
  • Dot Blot - Anti-Smad2 (phospho S250) antibody [EPR26263-58] (AB300079)
  • Anti-Smad2 (phospho S250) antibody [EPR26263-58] (AB300079)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EPR26263-58] to Smad2 (phospho S250)
  • Suitable for: WB, Dot blot, IP, ICC/IF
  • Knockout validated
  • Reacts with: Mouse, Rat, Human

You may also be interested in

Western blot
Product image
InstantBlue® Coomassie Protein Stain (ISB1L) (ab119211)
Conjugation
Product image
Alexa Fluor® 488 Conjugation Kit (Fast) - Lightning-Link® (ab236553)
Primary
Product image
Anti-c-Myc antibody [Y69] - ChIP Grade (ab32072)

View more associated products

Overview

  • Product name

    Anti-Smad2 (phospho S250) antibody [EPR26263-58]
    See all Smad2 primary antibodies
  • Description

    Rabbit monoclonal [EPR26263-58] to Smad2 (phospho S250)
  • Host species

    Rabbit
  • Tested applications

    Suitable for: WB, Dot blot, IP, ICC/IFmore details
    Unsuitable for: Flow Cyt (Intra) or IHC-P
  • Species reactivity

    Reacts with: Mouse, Rat, Human
  • Immunogen

    This product was produced with the following immunogens:
    Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.

    Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.

  • Positive control

    • WB: Whole cell lysates: Parental HeLa (human cervix adenocarcinoma epithelial cell) untreated and treated with 0.2µM PMA for 0.5h, NIH/3T3 untreated and treated with 0.2µM PMA for 0.5h, Untreated and treated with 0.2µM PMA for 0.5h C6 (rat glial tumor glial cell), HeLa treated with 0.2µM PMA for 0.5h (Untreated membrane ). ICC/IF: HeLa IP: HeLa and HIH/3T3 treated with 0.2µM PMA for 0.5h. DB: Smad2 peptides phosphorylated at: S245+S250+S255 a, S245+S250+S255 b, S245+S250, S250+S255, S250 only.
  • 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: 7.20
    Preservative: 0.01% Sodium azide
    Constituents: 59% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA
  • Concentration information loading...
  • Purity

    Protein A purified
  • Clonality

    Monoclonal
  • Clone number

    EPR26263-58
  • 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 S250) antibody [EPR26263-58] (BSA and Azide free) (ab300080)
  • Related Products

    • VeriBlot for IP Detection Reagent (HRP) (ab131366)
    • Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed (ab150081)
    • Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730)
    • Alexa Fluor® 594 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (ab195889)
    • Goat Anti-Rabbit IgG H&L (HRP) (ab97051)

Applications

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab300079 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
1/1000. Detects a band of approximately 65 kDa (predicted molecular weight: 52 kDa).
Dot blot
1/1000.
IP
1/30.
ICC/IF
1/50.
Notes
WB
1/1000. Detects a band of approximately 65 kDa (predicted molecular weight: 52 kDa).
Dot blot
1/1000.
IP
1/30.
ICC/IF
1/50.
Application notes
Is unsuitable for Flow Cyt (Intra) or IHC-P.

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 S250) antibody [EPR26263-58] (AB300079)
    Western blot - Anti-Smad2 (phospho S250) antibody [EPR26263-58] (AB300079)
    All lanes : Anti-Smad2 (phospho S250) antibody [EPR26263-58] (ab300079) at 1/1000 dilution

    Lane 1 : Untreated parental HeLa (human cervix adenocarcinoma epithelial cell) whole cell lysate
    Lane 2 : Parental HeLa treated with 0.2µM PMA for 0.5h whole cell lysate
    Lane 3 : Untreated SAMD2 KO Hela whole cell lysate
    Lane 4 : SAMD2 KO Hela treated with 0.2µM PMA for 0.5h whole cell lysate

    Lysates/proteins at 30 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution

    Predicted band size: 52 kDa
    Observed band size: 62 kDa why is the actual band size different from the predicted?


    Exposure time: 10 seconds


    5% NFDM/TBST was used as blocking and diluting buffer. 

  • Western blot - Anti-Smad2 (phospho S250) antibody [EPR26263-58] (AB300079)
    Western blot - Anti-Smad2 (phospho S250) antibody [EPR26263-58] (AB300079)
    All lanes : Anti-Smad2 (phospho S250) antibody [EPR26263-58] (ab300079) at 1/1000 dilution

    Lane 1 : Untreated NIH/3T3 (Mouse embryonic fibroblast) whole cell lysate
    Lane 2 : NIH/3T3 treated with 0.2µM PMA for 0.5h whole cell lysate

    Lysates/proteins at 20 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution

    Predicted band size: 52 kDa
    Observed band size: 65 kDa why is the actual band size different from the predicted?


    Exposure time: 180 seconds


    5% NFDM/TBST was used as blocking and diluting buffer. 

  • Western blot - Anti-Smad2 (phospho S250) antibody [EPR26263-58] (AB300079)
    Western blot - Anti-Smad2 (phospho S250) antibody [EPR26263-58] (AB300079)
    All lanes : Anti-Smad2 (phospho S250) antibody [EPR26263-58] (ab300079) at 1/1000 dilution

    Lane 1 : Untreated C6 (rat glial tumor cell) whole cell lysate
    Lane 2 : C6 treated with 0.2µM PMA for 0.5h whole cell lysate

    Lysates/proteins at 20 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution

    Predicted band size: 52 kDa
    Observed band size: 65 kDa why is the actual band size different from the predicted?


    Exposure time: 180 seconds


    5% NFDM/TBST was used as blocking and diluting buffer. 

    Bands around and below the 50-kDa marker could be degradation fragments.

  • Western blot - Anti-Smad2 (phospho S250) antibody [EPR26263-58] (AB300079)
    Western blot - Anti-Smad2 (phospho S250) antibody [EPR26263-58] (AB300079)
    All lanes : Anti-Smad2 (phospho S250) antibody [EPR26263-58] (ab300079) at 1/1000 dilution

    Lane 1 : HeLa (human cervix adenocarcinoma epithelial cell) treated with 0.2µM PMA for 0.5h (Untreated membrane) whole cell lysate
    Lane 2 : HeLa treated with 0.2µM PMA for 0.5h (phosphatase treated membrane) whole cell lysate

    Lysates/proteins at 20 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution

    Predicted band size: 52 kDa
    Observed band size: 65 kDa why is the actual band size different from the predicted?


    Exposure time: 180 seconds


    5% NFDM/TBST was used as blocking and diluting buffer. 

  • Immunocytochemistry/ Immunofluorescence - Anti-Smad2 (phospho S250) antibody [EPR26263-58] (AB300079)
    Immunocytochemistry/ Immunofluorescence - Anti-Smad2 (phospho S250) antibody [EPR26263-58] (AB300079)

    Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized HeLa (human cervix adenocarcinoma epithelial cell) cells lebeling Smad2 (phospho S250) with ab300079 at 1/50 (10.8 µg/ml) dilution, followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 (2 µg/mL) dilution (Green). Confocal image showing increased cytoplasm and nuclear staining in HeLa cells treated with PMA (200 nM ) for 30 min. ab195889 Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor® 594) was used to counterstain tubulin at 1/200 (2.5 µg/ml) dilution (Red). The Nuclear counterstain was DAPI (Blue).

    Secondary antibody only control: Secondary antibody is ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 (2 µg/mL) dilution.

  • Immunoprecipitation - Anti-Smad2 (phospho S250) antibody [EPR26263-58] (AB300079)
    Immunoprecipitation - Anti-Smad2 (phospho S250) antibody [EPR26263-58] (AB300079)

    Smad2 (phospho S250) was immunoprecipitated from 0.35 mg HeLa (human cervix adenocarcinoma epithelial cell) treated with 0.2µM PMA for 0.5h whole cell lysate with ab300079 at 1/30 dilution (2 µg in 0.35 mg lysates). Western blot was performed on the immunoprecipitate using ab300079 at 1/1000 dilution. VeriBlot for IP secondary antibody (HRP)(ab131366) was used at 1/5000 dilution.

    Lane 1: HeLa (human cervix adenocarcinoma epithelial cell) treated with 0.2µM PMA for 0.5h whole cell lysate 10 µg (Inset)

    Lane 2: ab300079 in HeLa treated with 0.2µM PMA for 0.5h whole cell lysate

    Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab300079 in HeLa treated with 0.2µM PMA for 0.5h whole cell lysate

    Blocking and dilution buffer and concentration: 5% NFDM/TBST.

    Exposure time: 10 seconds

     

  • Immunoprecipitation - Anti-Smad2 (phospho S250) antibody [EPR26263-58] (AB300079)
    Immunoprecipitation - Anti-Smad2 (phospho S250) antibody [EPR26263-58] (AB300079)

    Smad2 (phospho S250) was immunoprecipitated from 0.35 mg NIH/3T3 treated with 0.2µM PMA for 0.5h whole cell lysate with ab300079 at 1/30 dilution (2µg in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab300079 at 1/1000 dilution. VeriBlot for IP secondary antibody (HRP)(ab131366) was used at 1/5000 dilution.

    Lane 1: NIH/3T3 treated with 0.2µM PMA for 0.5h whole cell lysate 20 µg (Inset)

    Lane 2: ab300079 in NIH/3T3 treated with 0.2µM PMA for 0.5h whole cell lysate

    Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab300079 in NIH/3T3 whole cell lysate

    Blocking and dilution buffer and concentration: 5% NFDM/TBST.

    Exposure time: 10 seconds

     

  • Dot Blot - Anti-Smad2 (phospho S250) antibody [EPR26263-58] (AB300079)
    Dot Blot - Anti-Smad2 (phospho S250) antibody [EPR26263-58] (AB300079)

    Dot blot analysis of Smad2 (phospho S250) using ab300079 at 1:1000 (0.54 µg/ml), followed by a Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (ab97051) at 1:100,000 dilution. 

    Lane 1: Smad2 (phospho S245+S250+S255) peptide a
    Lane 2: Smad2 (phospho S245+S250+S255) peptide b
    Lane 3: Smad2 (phospho S245+S250) peptide
    Lane 4: Smad2 (phospho S245+S255) peptide
    Lane 5: Smad2 (phospho S250+S255) peptide
    Lane 6: Smad2 (phospho S245) peptide

    Lane 7: Smad2 (phospho S250) peptide
    Lane 8: Smad2 (phospho S255) peptide
    Lane 9: Smad2 non-phospho peptide

    Exposure time: 3 minutes

    Blocking and diluting buffer and concentration: 5% NFDM/TBST

  • Anti-Smad2 (phospho S250) antibody [EPR26263-58] (AB300079)
    Anti-Smad2 (phospho S250) antibody [EPR26263-58] (AB300079)

Protocols

To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.

Click here to view the general protocols

Datasheets and documents

  • SDS download

  • Datasheet download

    Download

Certificate of Compliance

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

References (0)

Publishing research using ab300079? Please let us know so that we can cite the reference in this datasheet.

ab300079 has not yet been referenced specifically in any publications.

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 ab300079.
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
© 1998-2023 Abcam plc. All rights reserved.