Smads are a family of intracellular proteins that transmit transforming growth factor beta (TGF-beta ) superfamily signals from the cell surface to the nucleus. The Smad family is divided into three subclasses: receptor regulated Smads, (Smads 1, 2, 3, 5 and 8); the common partner, (Smad4); and the inhibitory Smads, (Smads 6 and 7). The binding of TGF-beta or activin to their cognate receptor induces phosphorylation of Smads 2 and 3. The activated Smads associate with the common-mediator subunit, Smad4, and the heteromeric complex translocates into the nucleus to initiate transcription. Smad3, also known as Mothers Against Decapentaplegic homolog 3 (MADH3), shares 83% amino acid identity with Smad2, also known as Mothers Against Decapentaplegic homolog 2 (MADH2). Human Smad2 has 99% identity to mouse and rat Smad2. Human Smad3 has 99% identity to mouse and rat Smad3.
Human phospho-Smad2/3 (S465/S467) Antibody (1074A)
R&D Systems | Catalog # MAB8935
Recombinant Monoclonal Antibody.
Scientific images may be resized, cropped, or adjusted for brightness or contrast for presentation purposes. These updates are limited to presentation and do not affect the underlying scientific interpretation of the data. Where available, additional source data may be provided upon request.
Key Product Details
Validated by
Biological Validation
Species Reactivity
Validated:
Human
Cited:
Human, Mouse
Applications
Validated:
Western Blot
Cited:
Western Blot, Immunocytochemistry, Chromatin Immunoprecipitation (ChIP)
Label
Unconjugated
Antibody Source
Recombinant Monoclonal Rabbit IgG Clone # 1074A
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Product Specifications
Immunogen
Phosphopeptide containing the human Smad2 S465/S467 site
Accession # NP_005892
Accession # NP_005892
Specificity
Detects human Smad2 and Smad3 when dually phosphorylated at S465 and S467 in Western blots.
Clonality
Monoclonal
Host
Rabbit
Isotype
IgG
Scientific Data Images for Human phospho-Smad2/3 (S465/S467) Antibody (1074A)
Detection of Human Phospho-Smad2/3 (S465/S467) by Western Blot.
Western blot shows lysates of HepG2 human hepatocellular carcinoma cell line untreated (-) or treated (+) with 10 ng/mL Recombinant Human TGF-beta 1 (Catalog # 240-B) for 30 minutes. PVDF membrane was probed with 2 µg/mL of Rabbit Anti-Human Phospho-Smad2/3 (S465/S467) Monoclonal Antibody (Catalog # MAB8935) followed by HRP-conjugated Anti-Rabbit IgG Secondary Antibody (Catalog # HAF008). A specific band was detected for Phospho-Smad2/3 (S465/S467) at approximately 68 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 2.
Detection of Phospho-Smad2/3 (S465/S467) by Western Blot
PAO-induced production of IL-33 in primary human airway epithelial cells. BEAS-2B cells were treated with PAO (0, 20, 50, 100 nM) for 24 h. (a, b) Immunoblot and quantitative analysis for the expression of IL-33. (c) Representative immunoblot (d) quantitative analysis for the expression of pSMAD2/3, SMAD2/3, and GAPDH. Experiments were conducted in triplicate. Data are presented as mean ± SEM, *P < 0.05, ANOVA, as compared control (vehicle) to PAO-treated cells. Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/40195488), licensed under a CC-BY license. Not internally tested by R&D Systems.Applications for Human phospho-Smad2/3 (S465/S467) Antibody (1074A)
Application
Recommended Usage
Western Blot
2 µg/mL
Sample: HepG2 human hepatocellular carcinoma cell line treated with Recombinant Human TGF‑ beta 1 (Catalog # 240-B)
Sample: HepG2 human hepatocellular carcinoma cell line treated with Recombinant Human TGF‑ beta 1 (Catalog # 240-B)
Reviewed Applications
Read 2 reviews rated 4.5 using MAB8935 in the following applications:
Formulation, Preparation, and Storage
Purification
Protein A or G purified from cell culture supernatant
Reconstitution
Reconstitute at 0.2 mg/mL in sterile PBS. For liquid material, refer to CoA for concentration.
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Formulation
Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. *Small pack size (SP) is supplied either lyophilized or as a 0.2 µm filtered solution in PBS.
Shipping
Lyophilized product is shipped at ambient temperature. Liquid small pack size (-SP) is shipped with polar packs. Upon receipt, store immediately at the temperature recommended below.
Stability & Storage
Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
- 12 months from date of receipt, -20 to -70 °C as supplied.
- 1 month, 2 to 8 °C under sterile conditions after reconstitution.
- 6 months, -20 to -70 °C under sterile conditions after reconstitution.
Calculators
Background: Smad2/3
Long Name
Mothers Against DPP Homolog 2/3
Alternate Names
hMAD-2, hSMAD2, JV18, JV18-1SMAD, mothers against DPP homolog 2 (Drosophila), MAD homolog 2, Mad protein homolog, Mad, mothers against decapentaplegic homolog 2, MADH2MGC34440, MADR2MGC22139, Mad-related protein 2, mother against DPP homolog 2, mothers against decapentaplegic homolog 2, Mothers against DPP homolog 2, Sma- and Mad-related protein 2, SMAD 2, SMAD family member 2MAD, mothers against decapentaplegic homolog 2 (Drosophila), SMAD, mothers against DPP homolog 2, Smad2
UniProt
Additional Smad2/3 Products
Product Documents for Human phospho-Smad2/3 (S465/S467) Antibody (1074A)
Certificate of Analysis
To download a Certificate of Analysis, please enter a lot or batch number in the search box below.
Note: Certificate of Analysis not available for kit components.
Product Specific Notices for Human phospho-Smad2/3 (S465/S467) Antibody (1074A)
For research use only
Citations for Human phospho-Smad2/3 (S465/S467) Antibody (1074A)
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Customer Reviews for Human phospho-Smad2/3 (S465/S467) Antibody (1074A) (2)
4.5 out of 5
2 Customer Ratings
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Application: Western BlotSample Tested: primary astrocytesSpecies: HumanVerified Customer | Posted 08/21/2024Western blot using lysates from human primary astrocytes 15min after stimulation with human TGF-beta. 120ug of protein have been loaded.Lysates from primary human astrocytes
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Application: Western BlotSample Tested: Mv1Lu mink lung epithelial cell lineSpecies: MinkVerified Customer | Posted 07/08/2020
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Protocols
Find general support by application which include: protocols, troubleshooting, illustrated assays, videos and webinars.
- Cellular Response to Hypoxia Protocols
- R&D Systems Quality Control Western Blot Protocol
- Troubleshooting Guide: Western Blot Figures
- Western Blot Conditions
- Western Blot Protocol
- Western Blot Protocol for Cell Lysates
- Western Blot Troubleshooting
- Western Blot Troubleshooting Guide
- View all Protocols, Troubleshooting, Illustrated assays and Webinars
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