Smad1 Antibody [Unconjugated]

Smad1 was detected in immersion fixed paraffin-embedded sections of human breast cancer tissue using Goat Anti-Human Smad1 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF2039) at 15 µg/mL overnight at 4 °C. Tissue was stained using the Anti-Goat HRP-DAB Cell & Tissue Staining Kit (brown; Catalog # CTS008) and counterstained with hematoxylin (blue). Specific labeling was localized to the nuclei of glandular epithelial cells. View our protocol for Chromogenic IHC Staining of Paraffin-embedded Tissue Sections.

Western blot shows lysates of HepG2 human hepatocellular carcinoma cell line, MBA-MB-468 human breast cancer cell line, and HT-29 human colon adenocarcinoma cell line. PVDF membrane was probed with 0.5 µg/mL of Goat Anti-Human Smad1 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF2039) followed by HRP-conjugated Anti-Goat IgG Secondary Antibody (Catalog # HAF109). A specific band was detected for Smad1 at approximately 63 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.

Smad1 was detected in immersion fixed paraffin-embedded sections of human breast array using Goat Anti-Human Smad1 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF2039) at 15 µg/mL overnight at 4 °C. Tissue was stained using the Anti-Goat HRP-DAB Cell & Tissue Staining Kit (brown; Catalog # CTS008) and counterstained with hematoxylin (blue). Lower panel shows a lack of labeling if primary antibodies are omitted and tissue is stained only with secondary antibody followed by incubation with detection reagents. View our protocol for Chromogenic IHC Staining of Paraffin-embedded Tissue Sections.

Simple Western lane view shows lysates of HepG2 human hepatocellular carcinoma cell line and MDA‑MB‑468 human breast cancer cell line, loaded at 0.2 mg/mL. A specific band was detected for Smad1 at approximately 66 kDa (as indicated) using 5 µg/mL of Goat Anti-Human Smad1 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF2039) followed by 1:50 dilution of HRP-conjugated Anti-Goat IgG Secondary Antibody (Catalog # HAF109). This experiment was conducted under reducing conditions and using the12-230 kDa separation system.

Up-regulation of BMP signaling in the Fstl1-/- ureter and kidney.(A) Western blots of pSmad1/5/8, Smad1, pAKT (Ser473), AKT, and beta -actin from E15.5 (left panel) and E16.5 (second panel) ureter protein, E18.5 kidney protein (third panels), and HEK293 cells treated with the conditioned media containing BMP4 (20 ng/ml) transfected either with the Fstl1 or pcDNA3.1 vector (Mock) for 30 min (right panels). (B) Quantitative real-time PCR of Bmp2 (n = 5, p = 0.35), Bmp4 (n = 4, p = 0.73), Bmp5 (n = 5, p = 0.63), Bmp7 (n = 5, p = 0.50), TGF-beta 1 (n = 5, p = 0.51), Alk3 (n = 6, p = 0.40), Alk6 (n = 5, p = 0.21), Alk5 (n = 4, p = 0.47), BmprII (n = 6, p = 0.73), ActrIIa (n = 5, p = 0.97), and ActrIIb (n = 5, p = 0.73) from E16.5 ureters. (C) Co-immunoprecipitation of Myc-Fstl1 and HA-tagged BMP/TGF-beta receptors in HEK293 cells. Myc-Fstl1 can be immunoprecipitated with the anti-c-Myc antibody. Note that HA-ALK6 was co-immunoprecipitated by the anti-c-Myc and detected by the anti-HA antibody (lane 4). Scale bar: 40µm. Image collected and cropped by CiteAb from the following publication (//dx.plos.org/10.1371/journal.pone.0032554), licensed under a CC-BY license. Not internally tested by R&D Systems.

• Reactivity: Hu
• Applications: WB, Simple Western, IHC
• Clonality: Polyclonal
• Host: Goat
• Conjugate: Unconjugated
• Concentration: LYOPH

Smad1 Antibody [Unconjugated] Summary

• Immunogen: E. coli-derived recombinant human Smad1
  Asn2-Met454
  Accession # Q15797
• Specificity: Detects human Smad1 in direct ELISAs and Western blots. In direct ELISAs, approximately 15% cross-reactivity with recombinant human (rh) Smad5 is observed, and less than 5% cross-reactivity rhSmad4 and rhSmad9 is observed.
• Source: N/A
• Isotype: IgG
• Clonality: Polyclonal
• Host: Goat
• Gene: SMAD1
• Purity Statement: Antigen Affinity-purified

Applications/Dilutions

• Dilutions:
  • Immunohistochemistry 5-15 ug/mL
  • Simple Western 5 ug/mL
  • Western Blot 0.5 ug/mL
• Application Notes: In Simple Western only 10-15 uL of the recommended dilution is used per data point.

Packaging, Storage & Formulations

• 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.
• Buffer: 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.
• Preservative: No Preservative
• Concentration: LYOPH
• Reconstitution Instructions: Reconstitute at 0.2 mg/mL in sterile PBS.

Notes

This product is produced by and ships from R&D Systems, Inc., a Bio-Techne brand.

Alternate Names for Smad1 Antibody [Unconjugated]

• BSP1
• BSP-1
• hSMAD1
• JV4-1SMAD, mothers against DPP homolog 1 (Drosophila)
• MAD homolog 1
• MADH1JV41
• MADR1MAD, mothers against decapentaplegic homolog 1 (Drosophila)
• Mad-related protein 1
• mothers against decapentaplegic homolog 1
• Mothers against DPP homolog 1
• SMAD family member 1SMAD 1
• SMAD, mothers against DPP homolog 1
• Smad1
• TGF-beta signaling protein 1
• transforming growth factor-beta signaling protein 1

Background

Smads are a family of intracellular proteins that transmit transforming growth factor beta (TGF-beta ) superfamily signals from the cell surface to the nucleus. Upon signaling by some BMP family members, Smad1 is phosphorylated resulting in its association with the common-mediator subunit, Smad4. This heteromeric complex then translocates into the nucleus to exert transcriptional comodulator activity.

Limitations

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Primary Antibodies are guaranteed for 1 year from date of receipt.

Publications for Smad1 Antibody (AF2039)(8)

Publications using AF2039

• E Dalmo, P Johansson, M Niklasson, I Gustavsson, S Nelander, B Westermark Growth inhibitory activity of bone morphogenetic protein 4 in human glioblastoma cell lines is heterogeneous and dependent on reduced SOX2 expression Mol. Cancer Res., 2020-03-31;0(0):. 2020-03-31 [PMID: 32234828] (Western Blot, Human)
• Toshio Doi, Tatsumi Moriya, Yui Fujita, Naoto Minagawa, Masaru Usami, Tomoko Sasaki et al. Urinary IgG4 and Smad1 Are Specific Biomarkers for Renal Structural and Functional Changes in Early Stages of Diabetic Nephropathy Diabetes 2018-05-01 [PMID: 29490904]
• Karystinou A, Roelofs AJ, Neve A et al. Yes-associated protein (YAP) is a negative regulator of chondrogenesis in mesenchymal stem cells. Arthritis Res Ther. 5/30/2015 [PMID: 26025096]
• Jiang D, Aguiar R MicroRNA-155 controls RB phosphorylation in normal and malignant B lymphocytes via the noncanonical TGF-beta1/SMAD5 signaling module. Blood, 2013-10-17;123(1):86-93. 2013-10-17 [PMID: 24136167] (Western Blot, Human)
• Shimamoto , Seiko, Kubota , Yasuo, Yamaguchi , Fuminori, Tokumitsu , Hiroshi, Kobayashi , Ryoji Ca2+/S100 proteins act as upstream regulators of the chaperone-associated ubiquitin ligase CHIP (C terminus of Hsc70-interacting protein). J Biol Chem, 2013-01-23;288(10):7158-68. 2013-01-23 [PMID: 23344957] (Western Blot, Human)
• Olivier Nolan-Stevaux, Wendy Zhong, Stacey Culp, Kathy Shaffer, Joseph Hoover, Dineli Wickramasinghe et al. Endoglin Requirement for BMP9 Signaling in Endothelial Cells Reveals New Mechanism of Action for Selective Anti-Endoglin Antibodies PLoS ONE 2012-12-27 [PMID: 23300529]
• Alborzinia H, Schmidt-Glenewinkel H, Ilkavets I, Breitkopf-Heinlein K, Cheng X, Hortschansky P, Dooley S, Wolfl S Quantitative kinetics analysis of BMP2 uptake into cells and its modulation by BMP antagonists. J Cell Sci, 2012-10-17;126(0):117-27. 2012-10-17 [PMID: 23077176] (Western Blot, Human)
• Xu J, Qi X, Gong J Fstl1 antagonizes BMP signaling and regulates ureter development. PLoS ONE, 2012-04-02;7(4):e32554. 2012-04-02 [PMID: 22485132] (IHC, Mouse)

Bioinformatics

• Gene Symbol: SMAD1
• Uniprot:
  • Q15797()