Recombinant Anti-SMAD1 + SMAD5 + SMAD9 (phospho S463 + S465 + S467) antibody [MMC-1-104-3] (ab92698)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [MMC-1-104-3] to Smad1 (phospho S463 + S465) + SMAD5 (phospho S463 + S465) + SMAD9 (phospho S465 + S467)
- Suitable for: Dot blot, WB, IHC-P
- Reacts with: Mouse, Rat, Human
Related conjugates and formulations
Overview
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Product name
Anti-SMAD1 + SMAD5 + SMAD9 (phospho S463 + S465 + S467) antibody [MMC-1-104-3] -
Description
Rabbit monoclonal [MMC-1-104-3] to Smad1 (phospho S463 + S465) + SMAD5 (phospho S463 + S465) + SMAD9 (phospho S465 + S467) -
Host species
Rabbit -
Specificity
This antibody may cross-react with Smad1 Phospho (pS463/465) and Smad9 Phospho (pS465/467).
Stimulation may be required to allow detection of the phosphorylated protein. Please see images below for recommended treatment conditions and positive controls.
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Tested applications
Suitable for: Dot blot, WB, IHC-Pmore details
Unsuitable for: Flow Cyt or ICC/IF -
Species reactivity
Reacts with: Mouse, Rat, Human -
Immunogen
Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.
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Positive control
- WB: HeLa cell lysate treated with BMP-4. IHC-P: Human breast carcinoma, colonic carcinoma, tonsil, hepatocellular carcinoma, cervical carcinoma and glioma tissues.
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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. Stable for 12 months at -20°C. -
Storage buffer
pH: 7.20
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA -
Concentration information loading...
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Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
MMC-1-104-3 -
Isotype
IgG -
Research areas
Associated products
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Alternative Versions
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Compatible Secondaries
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Isotype control
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Recombinant Protein
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab92698 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 |
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Dot blot |
1/1000.
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WB | (1) |
1/1000 - 1/10000. Predicted molecular weight: 52 kDa.
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IHC-P |
1/100 - 1/800. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
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Notes |
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Dot blot
1/1000. |
WB
1/1000 - 1/10000. Predicted molecular weight: 52 kDa. |
IHC-P
1/100 - 1/800. Perform heat mediated antigen retrieval before commencing with IHC staining protocol. |
Target
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Cellular localization
Smad1: Cytoplasm. Nucleus. Cytoplasmic in the absence of ligand. Migrates to the nucleus when complexed with SMAD4. Co-localizes with LEMD3 at the nucleus inner membrane. SMAD5: Cytoplasm. Nucleus. Cytoplasmic in the absence of ligand. Migrates to the nucleus when complexed with SMAD4. SMAD9: Cytoplasm. Nucleus. In the cytoplasm in the absence of ligand. Migration to the nucleus when complexed with SMAD4. -
Database links
- Entrez Gene: 4086 Human
- Entrez Gene: 4090 Human
- Entrez Gene: 4093 Human
- Entrez Gene: 17125 Mouse
- Entrez Gene: 17129 Mouse
- Entrez Gene: 55994 Mouse
- Entrez Gene: 25671 Rat
- Entrez Gene: 59328 Rat
see all
Images
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All lanes : Anti-SMAD1 + SMAD5 + SMAD9 (phospho S463 + S465 + S467) antibody [MMC-1-104-3] (ab92698) at 1/5000 dilution (purified)
Lane 1 : Mouse brain tissue lysate
Lane 2 : Rat brain tissue lysate
Lysates/proteins at 10 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: 58 kDa why is the actual band size different from the predicted?Blocking and dilution buffer: 5% NFDM /TBST.
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All lanes : Anti-SMAD1 + SMAD5 + SMAD9 (phospho S463 + S465 + S467) antibody [MMC-1-104-3] (ab92698) at 1/10000 dilution (purified)
Lane 1 : HeLa whole cell lysate - untreated
Lane 2 : HeLa whole cell lysate - treated with Bone Morphogenetic protein-4
Lysates/proteins at 10 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/10000 dilution
Predicted band size: 52 kDa
Observed band size: 58 kDa why is the actual band size different from the predicted?Blocking and dilution buffer: 5% NFDM /TBST.
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of rat liver tissue labelling SMAD5 (phospho S463 + S465) with purified ab92698 at a dilution of 1/800. Heat mediated antigen retrieval was performed using Tris/EDTA buffer pH 9. ab97051, a HRP-conjugated goat anti-rabbit IgG (H+L) was used as the secondary antibody (1/500). Negative control using PBS instead of primary antibody. Counterstained with hematoxylin.
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of mouse liver tissue labelling SMAD5 (phospho S463 + S465) with purified ab92698 at a dilution of 1/800. Heat mediated antigen retrieval was performed using Tris/EDTA buffer pH 9. ab97051, a HRP-conjugated goat anti-rabbit IgG (H+L) was used as the secondary antibody (1/500). Negative control using PBS instead of primary antibody. Counterstained with hematoxylin.
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human cervical carcinoma tissue labelling SMAD5 (phospho S463 + S465) with purified ab92698 at a dilution of 1/800. Heat mediated antigen retrieval was performed using Tris/EDTA buffer pH 9. ab97051, a HRP-conjugated goat anti-rabbit IgG (H+L) was used as the secondary antibody (1/500). Negative control using PBS instead of primary antibody. Counterstained with hematoxylin.
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Dot blot analysis of SMAD5 (pS463 + pS465) peptide (Lane 1), SMAD5 (pS465) peptide (Lane 2), SMAD5 (pS463) peptide (Lane 3) and SMAD5 non-phospho peptide (Lane 4) labelling SMAD5 (pS465) with purified ab92698 at a dilution of 1/1000. ab97051 (Peroxidase conjugated goat anti-rabbit IgG (H+L)) was used as the secondary antibody at a dilution of 1/100000.
Blocking and dilution buffer: 5% NFDM/TBST.
Exposure time: 3 minutes.
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All lanes : Anti-SMAD1 + SMAD5 + SMAD9 (phospho S463 + S465 + S467) antibody [MMC-1-104-3] (ab92698) at 1/10000 dilution
Lane 1 : Untreated HeLa (human cervix adenocarcinoma) whole cell lysates 20µg
Lane 2 : HeLa (human cervix adenocarcinoma) treated with Bone Morphogenetic Protein-4 (BMP-4) whole cell lysates 20µg
Lane 3 : HeLa (human cervix adenocarcinoma) treated with Bone Morphogenetic Protein-4 (BMP-4) and phosphatase, whole cell lysates 20µg.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution
Predicted band size: 52 kDa
Observed band size: 58 kDa why is the actual band size different from the predicted?
Exposure time: 30 secondsBlocking and dilution buffer: 5% NFDM/TBST.
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of (1) human breast carcinoma and (2) human colonic carcinoma tissues labelling SMAD5 (phospho S463 + P465) with unpurified ab92698 at a dilution of 1/100.
Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of normal human tonsil tissue labelling SMAD5 (phospho S463 + S465) with unpurified ab92698.
Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human hepatocellular carcinoma tissue labelling SMAD5 (phospho S463 + S465) with unpurified ab92698.
Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human cervical carcinoma tissue labelling SMAD5 (phospho S463 + S465) with unpurified ab92698.
Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human glioma tissue labelling SMAD5 (phospho S463 + S465) with unpurified ab92698.
Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
Protocols
Datasheets and documents
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SDS download
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Datasheet download
References (42)
ab92698 has been referenced in 42 publications.
- Tang C et al. Omentin‑1 induces osteoblast viability and differentiation via the TGF‑β/Smad signaling pathway in osteoporosis. Mol Med Rep 25:N/A (2022). PubMed: 35179221
- Mead TJ et al. Proteolysis of fibrillin-2 microfibrils is essential for normal skeletal development. Elife 11:N/A (2022). PubMed: 35503090
- Zhang X et al. Effects of icariin on the fracture healing in young and old rats and its mechanism. Pharm Biol 59:1245-1255 (2021). PubMed: 34511043
- Kato T et al. Candesartan prevents arteriopathy progression in cerebral autosomal recessive arteriopathy with subcortical infarcts and leukoencephalopathy model. J Clin Invest 131:N/A (2021). PubMed: 34779414
- Zhang XY et al. Icariin regulates miR-23a-3p-mediated osteogenic differentiation of BMSCs via BMP-2/Smad5/Runx2 and WNT/β-catenin pathways in osteonecrosis of the femoral head. Saudi Pharm J 29:1405-1415 (2021). PubMed: 35002378