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.
We are constantly working hard to ensure we provide our customers with best in class antibodies. As a result of this work we are pleased to now offer this antibody in purified format. We are in the process of updating our datasheets. The purified format is designated 'PUR' on our product labels. If you have any questions regarding this update, please contact our Scientific Support team.
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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Western blot - Anti-SMAD1 + SMAD5 + SMAD9 (phospho S463 + S465 + S467) antibody [MMC-1-104-3] (ab92698)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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Western blot - Anti-SMAD1 + SMAD5 + SMAD9 (phospho S463 + S465 + S467) antibody [MMC-1-104-3] (ab92698)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) - Anti-SMAD1 + SMAD5 + SMAD9 (phospho S463 + S465 + S467) antibody [MMC-1-104-3] (ab92698)
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) - Anti-SMAD1 + SMAD5 + SMAD9 (phospho S463 + S465 + S467) antibody [MMC-1-104-3] (ab92698)
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) - Anti-SMAD1 + SMAD5 + SMAD9 (phospho S463 + S465 + S467) antibody [MMC-1-104-3] (ab92698)
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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Western blot - Anti-SMAD1 + SMAD5 + SMAD9 (phospho S463 + S465 + S467) antibody [MMC-1-104-3] (ab92698)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) - Anti-SMAD1 + SMAD5 + SMAD9 (phospho S463 + S465 + S467) antibody [MMC-1-104-3] (ab92698)
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) - Anti-SMAD1 + SMAD5 + SMAD9 (phospho S463 + S465 + S467) antibody [MMC-1-104-3] (ab92698)
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) - Anti-SMAD1 + SMAD5 + SMAD9 (phospho S463 + S465 + S467) antibody [MMC-1-104-3] (ab92698)
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) - Anti-SMAD1 + SMAD5 + SMAD9 (phospho S463 + S465 + S467) antibody [MMC-1-104-3] (ab92698)
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) - Anti-SMAD1 + SMAD5 + SMAD9 (phospho S463 + S465 + S467) antibody [MMC-1-104-3] (ab92698)
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