![Western Blot: Smad3 [p Ser423, p Ser425] Antibody [NBP1-77836] - Shows detection of endogenous Smad3 in stimulated cell lysates. Lysates were prepared from control cells (- lanes), or cells stimulated with 2 ng/ml TGF (+lanes) for 1 hour. This reagent recognizes phosphorylated Smad3 and has negligible reactivity against non-phosphorylated Smad3 protein. Personal Communication. YIgG Zhang, NIH, CCR, Bethesda, MD.](http://images.novusbio.com/fullsize/Smad3-[p-Ser423--p-Ser425]-Antibody-Western-Blot-NBP1-77836-img0005.jpg "Western Blot: Smad3 [p Ser423, p Ser425] Antibody [NBP1-77836] - Shows detection of endogenous Smad3 in stimulated cell lysates. Lysates were prepared from control cells (- lanes), or cells stimulated with 2 ng/ml TGF (+lanes) for 1 hour. This reagent recognizes phosphorylated Smad3 and has negligible reactivity against non-phosphorylated Smad3 protein. Personal Communication. YIgG Zhang, NIH, CCR, Bethesda, MD.")
| Reactivity | Hu, MuSpecies Glossary |
| Applications | WB, ELISA, Flow, ICC/IF, IHC, Single-Cell Western |
| Clonality | Polyclonal |
| Host | Rabbit |
| Conjugate | Unconjugated |
| Description | This affinity-purified antibody is directed against the phosphorylated form of human Smad3 protein at the [p Ser423] and [p Ser425] residues. The product was affinity purified from monospecific antiserum by immunoaffinity purification. Antiserum was first purified against the phosphorylated form of the immunizing peptide. The resultant affinity purified antibody was then cross adsorbed against the non-phosphorylated form of the immunizing peptide Store vial at -20C prior to opening. Aliquot contents and freeze at -20C or below for extended storage. Avoid cycles of freezing and thawing. Centrifuge product if not completely clear after standing at room temperature. This product is stable for several weeks at 4C as an undiluted liquid. Dilute only prior to immediate use. |
| Immunogen | Smad3 [p Ser423, p Ser425] Antibody was prepared from whole rabbit serum produced by repeated immunizations with a dual phosphorylated synthetic peptide corresponding to a c-terminal region with Serine 423 and Serine 425 of human Smad3 protein. (Uniprot: P84022) |
| Modification | p Ser423, p Ser425 |
| Specificity | This affinity-purified antibody is directed against the phosphorylated form of human Smad3 protein at the pS423 and pS425 residues. The resultant affinity purified antibody was then cross adsorbed against the non-phosphorylated form of the immunizing peptide. Reactivity occurs against human Smad3 pS423 and pS425 protein and the antibody is specific for the phosphorylated form of the protein. Reactivity with non-phosphorylated human Smad3 is minimal by ELISA and western blot. Expect reactivity against phosphorylated Smad1 and Smad5. Negligible reactivity is seen against other phosphorylated Smad family members. A BLAST analysis was used to suggest cross reactivity with Smad3 from human, Xenopus laevis, Xenopus tropicalis, zebrafish, rat, mouse, swine, bovine and chicken based on 100% sequence homology with the immunogen. Reactivity against homologues from other sources is not known. |
| Isotype | IgG |
| Clonality | Polyclonal |
| Host | Rabbit |
| Gene | SMAD3 |
| Purity | Immunogen affinity purified |
| Innovator's Reward | Test in a species/application not listed above to receive a full credit towards a future purchase. |
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| Application Notes | This affinity purified antibody has been tested for use in ELISA, immunohistochemistry, and western blot. Specific conditions for reactivity should be optimized by the end user. Expect a band approximately 48 kDa in size corresponding to phosphorylated Smad3 protein by western blotting in the appropriate stimulated tissue or cell lysate or extract. Less than 0.2% reactivity is observed against the non-phosphorylated form of the immunizing peptide. This antibody is phospho specific for dual phosphorylated pS423 and pS425 of Smad3. Stimulation with 2 ng/ml TGF-beta for 1 hour is suggested. |
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| Storage | Store at -20C short term. Aliquot and store at -80C long term. Avoid freeze-thaw cycles. |
| Buffer | 0.02 M Potassium Phosphate, 0.15 M Sodium Chloride, pH 7.2 |
| Preservative | 0.01% Sodium Azide |
| Purity | Immunogen affinity purified |
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![Smad3 [p Ser423, p Ser425] Antibody](/sites/all/modules/enterprise-tech/et_datasheets/images/novus_guarantee.png "Smad3 [p Ser423, p Ser425] Antibody")
| Images | Ratings | Applications | Species | Date | Details | ||||||||
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reviewed by:
Verified Customer |
IHC-Fr | Feline | 08/15/2017 |
Summary
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Secondary Antibodies |
Isotype Controls |
Research Areas for Smad3 Antibody (NBP1-77836)Find related products by research area.
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![Immunocytochemistry/Immunofluorescence: Smad3 [p Ser423, p Ser425] Antibody [NBP1-77836] - BH4 precursor, sepiapterin, normalizes NO level and ameliorates malignant phenotype of precancerous and cancerous cells. Representative images of the progression series cultivated in 3D ECM with or without sepiapterin (20 uM) and stained for phospho-SMAD3 or ERBB2. Nuclei were counterstained with DAPI (blue). Scale bars: 20 um. (Bottom) Quantification of phospho-SMAD3 (left) and ERBB2 levels (right) of the progression series. Error bars: mean +/- STDEV. *p < 0.05; **p < 0.01 and ***p < 0.001. Note that the effects of sepiapterin are only discernible in precancerous and cancerous cells, but not in non-malignant cells. (See the results of different concentrations of sepiapterin in Supplementary Fig. 8). Image collected and cropped by CiteAb from the following publication (www.nature.com/articles/s41598-019-43239-x) licensed under a CC-BY license.](http://images.novusbio.com/fullsize/Smad3-[p-Ser423--p-Ser425]-Antibody-Immunocytochemistry-Immunofluorescence-NBP1-77836-img0007.jpg "Immunocytochemistry/Immunofluorescence: Smad3 [p Ser423, p Ser425] Antibody [NBP1-77836] - BH4 precursor, sepiapterin, normalizes NO level and ameliorates malignant phenotype of precancerous and cancerous cells. Representative images of the progression series cultivated in 3D ECM with or without sepiapterin (20 uM) and stained for phospho-SMAD3 or ERBB2. Nuclei were counterstained with DAPI (blue). Scale bars: 20 um. (Bottom) Quantification of phospho-SMAD3 (left) and ERBB2 levels (right) of the progression series. Error bars: mean +/- STDEV. *p < 0.05; **p < 0.01 and ***p < 0.001. Note that the effects of sepiapterin are only discernible in precancerous and cancerous cells, but not in non-malignant cells. (See the results of different concentrations of sepiapterin in Supplementary Fig. 8). Image collected and cropped by CiteAb from the following publication (www.nature.com/articles/s41598-019-43239-x) licensed under a CC-BY license.")
![Immunohistochemistry: Smad3 [p Ser423, p Ser425] Antibody [NBP1-77836] - Used at 2.5 ug/ml to detect signal in a variety of tissues including multi-human, multi-brain and multi-cancer slides. This image shows strong nuclear staining in the majority of epidermal keratinocytes at 40X. Tissue was formalin-fixed and paraffin embedded. The image shows localization of the antibody as the precipitated red signal, with a hematoxylin purple nuclear counterstain. Personal Communication, Tina Roush, LifeSpanBiosciences, Seattle, WA.](http://images.novusbio.com/fullsize/Smad3-[p-Ser423--p-Ser425]-Antibody-Immunohistochemistry-NBP1-77836-img0006.jpg "Immunohistochemistry: Smad3 [p Ser423, p Ser425] Antibody [NBP1-77836] - Used at 2.5 ug/ml to detect signal in a variety of tissues including multi-human, multi-brain and multi-cancer slides. This image shows strong nuclear staining in the majority of epidermal keratinocytes at 40X. Tissue was formalin-fixed and paraffin embedded. The image shows localization of the antibody as the precipitated red signal, with a hematoxylin purple nuclear counterstain. Personal Communication, Tina Roush, LifeSpanBiosciences, Seattle, WA.")
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![Western Blot: Goat anti-Rabbit IgG (H+L) Secondary Antibody [HRP] [NB7160] - Western blot showing vemurafenib treatment in BRAFV600E CRC cells inhibits fission mediator DRP1 with no significant effect on fusion proteins (Mfn1 & 2) using MFN-1 antibody (NBP1-51841) and corresponding secondary antibody, goat anti-rabbit IgG-HRP (NB7160). Image collected and cropped by CiteAb from the following publication (https://pubmed.ncbi.nlm.nih.gov/33738242).](https://images.novusbio.com/images/Goat-anti-Rabbit-IgG-H+L-Secondary-Antibody-HRP-Western-Blot-NB7160-img0001.jpg "Western Blot: Goat anti-Rabbit IgG (H+L) Secondary Antibody [HRP] [NB7160] - Western blot showing vemurafenib treatment in BRAFV600E CRC cells inhibits fission mediator DRP1 with no significant effect on fusion proteins (Mfn1 & 2) using MFN-1 antibody (NBP1-51841) and corresponding secondary antibody, goat anti-rabbit IgG-HRP (NB7160). Image collected and cropped by CiteAb from the following publication (https://pubmed.ncbi.nlm.nih.gov/33738242).")
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