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--smad3-antibody-epr19557-4-chip-grade-ab202445.pdf

  1. Send me a copy of this email
    I agree to the terms and conditions.
Signal Transduction Growth Factors/Hormones TGF
Share by email
RecombinantRabMAb

Recombinant Anti-Smad2 + Smad3 antibody [EPR19557-4] - ChIP Grade (ab202445)

  • Datasheet
  • SDS
  • Certificate of Compliance
Reviews (1) Submit a question References (26)

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 + Smad3 antibody [EPR19557-4] - ChIP Grade (ab202445)
  • Western blot - Anti-Smad2 + Smad3 antibody [EPR19557-4] - ChIP Grade (ab202445)
  • Western blot - Anti-Smad2 + Smad3 antibody [EPR19557-4] - ChIP Grade (ab202445)
  • Western blot - Anti-Smad2 + Smad3 antibody [EPR19557-4] - ChIP Grade (ab202445)
  • Immunocytochemistry/ Immunofluorescence - Anti-Smad2 + Smad3 antibody [EPR19557-4] - ChIP Grade (ab202445)
  • Flow Cytometry (Intracellular) - Anti-Smad2 + Smad3 antibody [EPR19557-4] - ChIP Grade (ab202445)
  • Immunoprecipitation - Anti-Smad2 + Smad3 antibody [EPR19557-4] - ChIP Grade (ab202445)
  • ChIP - Anti-Smad2 + Smad3 antibody [EPR19557-4] - ChIP Grade (ab202445)
  • Anti-Smad2 + Smad3 antibody [EPR19557-4] - ChIP Grade (ab202445)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EPR19557-4] to Smad2 + Smad3 - ChIP Grade
  • Suitable for: Flow Cyt (Intra), WB, ICC/IF, IP, ChIP
  • Reacts with: Mouse, Rat, Human

Conjugates logo Related conjugates and formulations

Carrier Free

You may also be interested in

Secondary
Product image
Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077)
Primary
Product image
Anti-IL-1 beta antibody [EPR21086] - BSA and Azide free (ab229696)
Knockout
Product image
Human SMAD2 knockout HeLa cell line (ab255430)

View more associated products

Overview

  • Product name

    Anti-Smad2 + Smad3 antibody [EPR19557-4] - ChIP Grade
    See all Smad2 + Smad3 primary antibodies
  • Description

    Rabbit monoclonal [EPR19557-4] to Smad2 + Smad3 - ChIP Grade
  • Host species

    Rabbit
  • Tested applications

    Suitable for: Flow Cyt (Intra), WB, ICC/IF, IP, ChIPmore details
  • Species reactivity

    Reacts with: Mouse, Rat, Human
  • Immunogen

    Recombinant fragment. This information is proprietary to Abcam and/or its suppliers.

  • Positive control

    • WB: Recombinant protein fragment human Smad2 and recombinant protein fragment human Smad3; HEK-293, HepG2, HeLa, Jurkat and C6 whole cell lysates; human fetal heart and fetal kidney lysates; mouse brain and heart lysates; rat brain and spleen lysates. ICC/IF: HeLa cells. Flow Cyt (intra): HeLa cells. IP: HeLa whole cell lysate. ChIP: Chromatin from HaCaT cells treated with 7ng/ml TGF-ß for 1h.
  • 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 (glycerin, glycerine), 0.05% BSA
  • Concentration information loading...
  • Purity

    Protein A purified
  • Clonality

    Monoclonal
  • Clone number

    EPR19557-4
  • Isotype

    IgG
  • Research areas

    • Signal Transduction
    • Growth Factors/Hormones
    • TGF
    • Signal Transduction
    • Signaling Pathway
    • Nuclear Signaling
    • SMADs
    • Epigenetics and Nuclear Signaling
    • Nuclear Signaling Pathways
    • SMADs
    • Stem Cells
    • Signaling Pathways
    • TGF beta
    • Cytoplasmic
    • Cancer
    • Signal transduction
    • Nuclear signaling
    • SMAD family
    • Epigenetics and Nuclear Signaling
    • ChIP assays
    • ChIP antibodies
    • Cancer
    • Cancer Metabolism
    • Response to hypoxia
    • Metabolism
    • Pathways and Processes
    • Metabolism processes
    • Hypoxia

Associated products

  • Alternative Versions

    • Anti-Smad2 + Smad3 antibody [EPR19557-4] - BSA and Azide free (ab232326)
  • ChIP Related Products

    • ChIP Kit (ab500)
  • Compatible Secondaries

    • Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077)
    • Goat Anti-Rabbit IgG H&L (HRP) (ab97051)
  • Conjugation kits

    • PE / R-Phycoerythrin Conjugation Kit - Lightning-Link® (ab102918)
    • APC Conjugation Kit - Lightning-Link® (ab201807)
  • Isotype control

    • Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730)
  • Related Products

    • VeriBlot for IP Detection Reagent (HRP) (ab131366)

Applications

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab202445 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
Flow Cyt (Intra)
1/600.
WB
1/1000. Detects a band of approximately 58-62 kDa (predicted molecular weight: 52 kDa).
ICC/IF
1/200.
IP
1/40.
ChIP
Use 2 µg for 25 µg of chromatin.
Notes
Flow Cyt (Intra)
1/600.
WB
1/1000. Detects a band of approximately 58-62 kDa (predicted molecular weight: 52 kDa).
ICC/IF
1/200.
IP
1/40.
ChIP
Use 2 µg for 25 µg of chromatin.

Target

  • Relevance

    SMAD is a family of proteins similar to the gene products of the Drosophila gene 'mothers against decapentaplegic' (Mad) and the C. elegans gene Sma. SMAD proteins are signal transducers and transcriptional modulators that mediate multiple signaling pathways. They mediate the signal of the transforming growth factor (TGF)-beta, and thus regulate multiple cellular processes, such as cell proliferation, apoptosis, and differentiation.
  • Cellular localization

    Cytoplasm. Nucleus. Note: Cytoplasmic in the absence of ligand. Migrates to the nucleus when complexed with SMAD4.
  • Database links

    • Entrez Gene: 4087 Human
    • Entrez Gene: 4088 Human
    • Entrez Gene: 17126 Mouse
    • Entrez Gene: 17127 Mouse
    • Entrez Gene: 25631 Rat
    • Entrez Gene: 29357 Rat
    • Omim: 601366 Human
    • Omim: 603109 Human
    • SwissProt: P84022 Human
    • SwissProt: Q15796 Human
    • SwissProt: Q62432 Mouse
    • SwissProt: Q8BUN5 Mouse
    • SwissProt: O70436 Rat
    • SwissProt: P84025 Rat
    see all
  • Alternative names

    • hMAD 2 antibody
    • hMAD 3 antibody
    • hSMAD2 antibody
    • hSMAD3 antibody
    • Mad related protein 2 antibody
    • MADH2 antibody
    • MADH3 antibody
    • MADR2 antibody
    • Mothers against DPP homolog 2 antibody
    • Mothers against DPP homolog 3 antibody
    • Sma and Mad related protein 2 antibody
    • SMA and MAD related protein 3 antibody
    • SMAD 2 antibody
    • SMAD 3 antibody
    • SMAD family member 2 antibody
    • SMAD family member 3 antibody
    • smad2+3 antibody
    • smad2/3 antibody
    see all

Images

  • Western blot - Anti-Smad2 + Smad3 antibody [EPR19557-4] - ChIP Grade (ab202445)
    Western blot - Anti-Smad2 + Smad3 antibody [EPR19557-4] - ChIP Grade (ab202445)
    All lanes : Anti-Smad2 + Smad3 antibody [EPR19557-4] - ChIP Grade (ab202445) at 1/1000 dilution

    Lane 1 : Recombinant protein fragment human Smad2
    Lane 2 : Recombinant protein fragment human Smad3

    Lysates/proteins at 0.01 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: 38,60 kDa why is the actual band size different from the predicted?


    Exposure time: 1 second


    Blocking/Dilution buffer: 5% NFDM/TBST.

    Human Smad2 fragment recombinant protein contains aa2-270 with His-Tag®. Human Smad3 fragment recombinant protein contains aa2-227 with His-Tag® and GST-tag.

  • Western blot - Anti-Smad2 + Smad3 antibody [EPR19557-4] - ChIP Grade (ab202445)
    Western blot - Anti-Smad2 + Smad3 antibody [EPR19557-4] - ChIP Grade (ab202445)
    All lanes : Anti-Smad2 + Smad3 antibody [EPR19557-4] - ChIP Grade (ab202445) at 1/2000 dilution

    Lane 1 : 293 (Human epithelial cell line from embryonic kidney) whole cell lysate
    Lane 2 : HepG2 (Human liver hepatocellular carcinoma cell line) whole cell lysate
    Lane 3 : HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate
    Lane 4 : Jurkat (Human T cell leukemia cell line from peripheral blood) whole cell lysate

    Lysates/proteins at 10 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: 58-62 kDa why is the actual band size different from the predicted?



    Blocking/Dilution buffer: 5% NFDM/TBST.

    Exposure time: Lane 1/2: 10 seconds; Lane 3: 3 seconds; Lane 4: 1 seconds.

  • Western blot - Anti-Smad2 + Smad3 antibody [EPR19557-4] - ChIP Grade (ab202445)
    Western blot - Anti-Smad2 + Smad3 antibody [EPR19557-4] - ChIP Grade (ab202445)
    All lanes : Anti-Smad2 + Smad3 antibody [EPR19557-4] - ChIP Grade (ab202445) at 1/2000 dilution

    Lane 1 : Human fetal heart lysate
    Lane 2 : Human fetal kidney lysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG Peroxidase Conjugate, specific to the non-reduced form of IgG at 1/10000 dilution

    Predicted band size: 52 kDa
    Observed band size: 58-62 kDa why is the actual band size different from the predicted?



    Blocking/Dilution buffer: 5% NFDM/TBST.

    Exposure time: Lane 1: 30 seconds; Lane 2: 10 seconds.

  • Western blot - Anti-Smad2 + Smad3 antibody [EPR19557-4] - ChIP Grade (ab202445)
    Western blot - Anti-Smad2 + Smad3 antibody [EPR19557-4] - ChIP Grade (ab202445)
    Lane 1 : Anti-Smad2 + Smad3 antibody [EPR19557-4] - ChIP Grade (ab202445) at 1/1000 dilution
    Lanes 2 & 5 : Anti-Smad2 + Smad3 antibody [EPR19557-4] - ChIP Grade (ab202445) at 1/2000 dilution
    Lanes 3-4 : Anti-Smad2 + Smad3 antibody [EPR19557-4] - ChIP Grade (ab202445) at 1/20000 dilution

    Lane 1 : Mouse brain lysate
    Lane 2 : Mouse heart lysate
    Lane 3 : Rat brain lysate
    Lane 4 : C6 (Rat glial tumor cell line) whole cell lysate
    Lane 5 : Rat spleen lysate

    Lysates/proteins at 10 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: 58-62 kDa why is the actual band size different from the predicted?



    Blocking/Dilution buffer: 5% NFDM/TBST.

    Exposure time: Lane 1: 3 minutes; Lane 2: 10 seconds; Lane 3: 30 seconds; Lane 4: 3 seconds; Lane 5: 5 seconds.

     

  • Immunocytochemistry/ Immunofluorescence - Anti-Smad2 + Smad3 antibody [EPR19557-4] - ChIP Grade (ab202445)
    Immunocytochemistry/ Immunofluorescence - Anti-Smad2 + Smad3 antibody [EPR19557-4] - ChIP Grade (ab202445)

    Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized HeLa (Human epithelial cell line from cervix adenocarcinoma) cells labeling Smad2 + Smad3 with ab202445 at 1/200 dilution, followed by Goat Anti-Rabbit IgG (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution (green). The results show signal translocation after TGF-beta (10ng/ml, 1h) treatment on HeLa cells.The nuclear counter stain is DAPI (blue).

    Tubulin is detected with Anti-alpha Tubulin mouse MAb (ab7291) at 1/1000 dilution followed by Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) (ab150120) secondary antibody at 1/1000 dilution (red).

    The negative controls are as follows:
    -ve control 1: ab202445 at 1/200 dilution, followed by Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) (ab150120) secondary antibody at 1/1000 dilution.
    -ve control 2: Anti-alpha Tubulin mouse MAb (ab7291) at 1/1000 dilution,followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution.

  • Flow Cytometry (Intracellular) - Anti-Smad2 + Smad3 antibody [EPR19557-4] - ChIP Grade (ab202445)
    Flow Cytometry (Intracellular) - Anti-Smad2 + Smad3 antibody [EPR19557-4] - ChIP Grade (ab202445)

    Intracellular flow cytometric analysis of 4% paraformaldehyde-fixed HeLa (Human epithelial cell line from cervix adenocarcinoma) cells labeling Smad2 + Smad3 with ab202445 at 1/600 dilution (red) compared with a rabbit monoclonal IgG isotype control (ab172730; black) and an unlabelled control (cells without incubation with primary antibody and secondary antibody; blue). Goat anti rabbit IgG (Alexa Fluorr® 488) at 1/2000 dilution was used as the secondary antibody. 

     

     

  • Immunoprecipitation - Anti-Smad2 + Smad3 antibody [EPR19557-4] - ChIP Grade (ab202445)
    Immunoprecipitation - Anti-Smad2 + Smad3 antibody [EPR19557-4] - ChIP Grade (ab202445)

    Smad2 + Smad3 was immunoprecipitated from 0.35mg of HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate with ab202445 at 1/40 dilution. Western blot was performed from the immunoprecipitate using ab202445 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/10000 dilution.

    Lane 1: HeLa whole cell lysate 10µg (Input).

    Lane 2: ab202445 IP in HeLa whole cell lysate.

    Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab202445 in HeLa whole cell lysate.

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

    Exposure time: 3 seconds.

  • ChIP - Anti-Smad2 + Smad3 antibody [EPR19557-4] - ChIP Grade (ab202445)
    ChIP - Anti-Smad2 + Smad3 antibody [EPR19557-4] - ChIP Grade (ab202445)

    Chromatin was prepared from HaCaT (Human keratinocyte cell line) cells treated with 7ng/ml TGF-β for 1h and non-treated according to the Abcam X-ChIP protocol. Cells were fixed with formaldehyde for 10 minutes. The ChIP was performed with 25µg of chromatin, 2µg of ab202445 (blue), and 20µl of Anti rabbit IgG sepharose beads. 2μg of rabbit normal IgG was added to the beads control (yellow). The immunoprecipitated DNA was quantified by real time PCR (Sybr green approach).

  • Anti-Smad2 + Smad3 antibody [EPR19557-4] - ChIP Grade (ab202445)
    Anti-Smad2 + Smad3 antibody [EPR19557-4] - ChIP Grade (ab202445)

Protocols

  • ChIP protocols
  • Flow cytometry protocols
  • Immunoprecipitation protocols
  • Immunocytochemistry & immunofluorescence protocols
  • Western blot protocols

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 (26)

Publishing research using ab202445? Please let us know so that we can cite the reference in this datasheet.

ab202445 has been referenced in 26 publications.

  • Xu J  et al. Uncarboxylated osteocalcin promotes proliferation and metastasis of MDA-MB-231 cells through TGF-ß/SMAD3 signaling pathway. BMC Mol Cell Biol 23:18 (2022). WB ; Human . PubMed: 35413833
  • Yang T  et al. Punicalin Alleviates OGD/R-Triggered Cell Injury via TGF-ß-Mediated Oxidative Stress and Cell Cycle in Neuroblastoma Cells SH-SY5Y. Evid Based Complement Alternat Med 2021:6671282 (2021). PubMed: 33628309
  • Liu L  et al. A high level of lncFGD5-AS1 inhibits epithelial-to-Mesenchymal transition by regulating the miR-196a-5p/SMAD6/BMP axis in gastric Cancer. BMC Cancer 21:453 (2021). PubMed: 33892661
  • Zhu Z  et al. NAT10 Promotes Osteogenic Differentiation of Mesenchymal Stem Cells by Mediating N4-Acetylcytidine Modification of Gremlin 1. Stem Cells Int 2021:8833527 (2021). PubMed: 33953754
  • Shen K  et al. Acetyl oxygen benzoate engeletin ester promotes KLF4 degradation leading to the attenuation of pulmonary fibrosis via inhibiting TGFß1-smad/p38MAPK-lnc865/lnc556-miR-29b-2-5p-STAT3 signal pathway. Aging (Albany NY) 13:13807-13821 (2021). PubMed: 33929970
View all Publications for this product

Customer reviews and Q&As

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

IHC - Wholemount abreview for Anti-Smad2 + Smad3 antibody [EPR19557-4] - ChIP Grade

Average
Abreviews
Abreviews
abreview image
Application
IHC - Wholemount
Sample
Mouse Tissue (Cornea)
Specification
Cornea
Read More
The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

Abcam user community

Verified customer

Submitted Nov 10 2020

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.