Mouse TGF-beta 2 Antibody

Detection of Human and Mouse TGF‑ beta2 by Western Blot.

Western blot shows lysates of Y-79 human retinoblastoma cell line, HUVEC human umbilical vein endothelial cells, NIH-3T3 mouse embryonic fibroblast cell line, 4T1 mouse breast cancer cell line, and NMuMG mouse mammary gland epithelial cell line. PVDF membrane was probed with 2 µg/mL of Rat Anti-Mouse TGF-beta 2 Monoclonal Antibody (Catalog # MAB73461) followed by HRP-conjugated Anti-Rat IgG Secondary Antibody (Catalog # HAF005). A specific band was detected for TGF-beta 2 at approximately 52 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.

TGF‑ beta2 in Mouse Embryo.

TGF-beta 2 was detected in immersion fixed frozen sections of mouse embryo (15 d.p.c.) using Rat Anti-Mouse TGF-beta 2 Monoclonal Antibody (Catalog # MAB73461) at 25 µg/mL overnight at 4 °C. Tissue was stained using the Anti-Rat HRP-DAB Cell & Tissue Staining Kit (brown; Catalog # CTS017) and counterstained with hematoxylin (blue). Specific staining was localized to neuronal processes. View our protocol for Chromogenic IHC Staining of Frozen Tissue Sections.

Detection of Human TGF-beta 2 by Western Blot

LOXL2 acts with TGF-beta through PI3K/ATK/mTORC1 signalling to control myofibroblast transformation and migration.(a) Western blot analysis of LOXL2, SMAD2, p-SMAD2 and GAPDH in human primary cardiac fibroblasts transfected with control (Ctrl) or LOXL2 siRNA. (b) Quantification of TGF-beta isoforms in the culture media of human primary cardiac fibroblasts transfected with control or LOXL2 siRNA (n=4). P value: Student's t-test. Error bar: s.e.m. (c) Changes of COLA1, alpha-SMA and FN1 mRNA in human primary cardiac fibroblasts transfected with control (Ctrl) or LOXL2 siRNA. (d) Western blot of LOXL2, p-AKT, AKT, p-S6K, S6K and p-4E-BP1 with or without PI3K inhibitors LY294002/PI828 (10 μM each), PI3K alpha inhibitors A66/BYL719 and mTORC1 inhibitor Rapamycine (0.1 μM) in cells infected with Ad_GFP/Ad_LOXL2. (e) TGF-beta 2 protein in the culture media (n=4). P value: Student's t-test. Error bar: s.e.m. (f,g) Western blot (f) and quantification (g) of TGF-beta 2 protein in the mice heart ventricles 6 weeks after sham/TAC operation. n=4 mice per group. P value: Student's t-test. Error bar: s.e.m. (h) A signalling cascade from LOXL2 to PI3K/AKT/mTORC1 to TGF-beta 2 translation. (i) Representative phase-contrast image of human primary cardiac fibroblasts 72 h after transfection with control (Ctrl) or LOXL2 siRNA. Scale bars, 50 μm. (j) Immunostaining of Col1A in IgG1- or alpha-LOXL2-treated mice 10 weeks (10W) after sham or TAC operation. Scale bars, 100 μm. Blue: haematoxylin. Brown: Col1A. (k,l) Gap closure assay of fibroblast migration in control, TGF-beta 2, alpha-LOXL2 (k) groups and quantification of cells migrating into the gap (l). Scale bars, 200 μm. n=5–10, P value: Student's t-test. Error bar: s.e.m. Image collected and cropped by CiteAb from the following publication (https://pubmed.ncbi.nlm.nih.gov/27966531), licensed under a CC-BY license. Not internally tested by R&D Systems.