Recombinant Anti-Smad2 + Smad3 antibody [EPR19557-4] - ChIP Grade (ab202445)
![Western blot - Anti-Smad2 + Smad3 antibody [EPR19557-4] - ChIP Grade (ab202445)](https://www.abcam.com/ps/products/202/ab202445/Images/ab202445-260125-anti-smad2-smad3-antibody-epr19557-4-chip-grade-western-blot.jpg "Western blot - 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
Related conjugates and formulations
Overview
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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.
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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.
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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
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
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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
Associated products
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Alternative Versions
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ChIP Related Products
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Compatible Secondaries
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Conjugation kits
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Isotype control
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Related Products
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.
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| WB |
1/1000. Detects a band of approximately 58-62 kDa (predicted molecular weight: 52 kDa).
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| ICC/IF |
1/200.
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| IP |
1/40.
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| ChIP |
Use 2 µg for 25 µg of chromatin.
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| Notes |
|---|
|
Flow Cyt (Intra)
1/600. |
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WB
1/1000. Detects a band of approximately 58-62 kDa (predicted molecular weight: 52 kDa). |
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ICC/IF
1/200. |
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IP
1/40. |
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ChIP
Use 2 µg for 25 µg of chromatin. |
Target
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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
see all -
Alternative names
- hMAD 2 antibody
- hMAD 3 antibody
- hSMAD2 antibody
see all
Images
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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 secondBlocking/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.
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![Western blot - Anti-Smad2 + Smad3 antibody [EPR19557-4] - ChIP Grade (ab202445)](https://www.abcam.com/ps/products/202/ab202445/Images/ab202445-260123-anti-smad2-smad3-antibody-epr19557-4-chip-grade-western-blot.jpg)
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.
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![Western blot - Anti-Smad2 + Smad3 antibody [EPR19557-4] - ChIP Grade (ab202445)](https://www.abcam.com/ps/products/202/ab202445/Images/ab202445-260120-anti-smad2-smad3-antibody-epr19557-4-chip-grade-western-blot.jpg)
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.
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![Western blot - Anti-Smad2 + Smad3 antibody [EPR19557-4] - ChIP Grade (ab202445)](https://www.abcam.com/ps/products/202/ab202445/Images/ab202445-260116-anti-smad2-smad3-antibody-epr19557-4-chip-grade-western-blot.jpg)
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.
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![Immunocytochemistry/ Immunofluorescence - Anti-Smad2 + Smad3 antibody [EPR19557-4] - ChIP Grade (ab202445)](https://www.abcam.com/ps/products/202/ab202445/Images/ab202445-260115-anti-smad2-smad3-antibody-epr19557-4-chip-grade-immunofluorescence.jpg)
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)](https://www.abcam.com/ps/products/202/ab202445/Images/ab202445-260114-anti-smad2-smad3-antibody-epr19557-4-chip-grade-flow-cytometry.jpg)
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.
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![Immunoprecipitation - Anti-Smad2 + Smad3 antibody [EPR19557-4] - ChIP Grade (ab202445)](https://www.abcam.com/ps/products/202/ab202445/Images/ab202445-260113-anti-smad2-smad3-antibody-epr19557-4-chip-grade-immunoprecipitation.jpg)
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.
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![ChIP - Anti-Smad2 + Smad3 antibody [EPR19557-4] - ChIP Grade (ab202445)](https://www.abcam.com/ps/products/202/ab202445/Images/ab202445-260131-anti-smad2-smad3-antibody-epr19557-4-chip-grade-chip.jpg)
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).
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![Anti-Smad2 + Smad3 antibody [EPR19557-4] - ChIP Grade (ab202445)](https://www.abcam.com/ps/products/202/ab202445/Images/ab202445-10-benefits-of-recombinant-antibodies.png)
Anti-Smad2 + Smad3 antibody [EPR19557-4] - ChIP Grade (ab202445)
Protocols
Datasheets and documents
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SDS download
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Datasheet download
Certificate of Compliance
References (26)
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