Human Smad1 Antibody

Smad1 in Human Breast Cancer Tissue. 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.

Detection of Human Smad1 by Western Blot. 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 in Human Breast. 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.

Detection of Human Smad1 by Simple Western^TM. 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 the 12-230 kDa separation system.

Detection of Mouse Smad1 by Western Blot 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 (https://dx.plos.org/10.1371/journal.pone.0032554), licensed under a CC-BY license. Not internally tested by R&D Systems.