|Detection of Human LAP (TGF-beta 1) by Western Blot. Western blot shows lysates of Saos‑2 human osteosarcoma cell line. PVDF membrane was probed with 2 µg/mL of Goat Anti-Human LAP (TGF-beta 1) Antigen Affinity-purified Polyclonal Antibody (Catalog # AF-246-NA) followed by HRP-conjugated Anti-Goat IgG Secondary Antibody (Catalog # HAF017). A specific band was detected for LAP (TGF-beta 1) at approximately 48 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.|
Detection of Human LAP (TGF-beta 1) by Simple WesternTM. Simple Western lane view shows lysates of K562 human chronic myelogenous leukemia cell line and A549 human lung carcinoma cell line, loaded at 0.2 mg/mL. A specific band was detected for LAP (TGF-beta 1) at approximately 61 kDa (as indicated) using 20 µg/mL of Goat Anti-Human LAP (TGF-beta 1) Antigen Affinity-purified Polyclonal Antibody (Catalog # AF-246-NA) 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.
|LAP TGF‑ beta 1 Inhibition of TGF‑ beta 1 Activity and Neutralization by Human LAP TGF‑ beta 1 Antibody. Recombinant Human LAP TGF‑ beta 1 (Catalog # 246‑LP) inhibits Recombinant Human TGF‑ beta 1 (Catalog # 240‑B) growth inhibition activity in the HT‑2 mouse T cell line in a dose-dependent manner (orange line). Inhibition of Recombinant Human TGF‑ beta 1 (1 ng/mL) activity elicited by Recombinant Human LAP TGF‑ beta 1 (500 ng/mL) is neutralized (green line) by increasing concentrations of Goat Anti-Human LAP TGF‑ beta 1 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF-246-NA). The ND50 is typically 0.4‑2 µg/mL.|
TGF-beta 1 (transforming growth factor beta 1) and the closely related TGF-beta 2 and -beta 3 are members of the large TGF-beta superfamily. TGF‑ beta proteins are highly pleiotropic cytokines that regulate processes such as immune function, proliferation and epithelial-mesenchymal transition (1‑3). Human TGF-beta 1 cDNA encodes a 390 amino acid (aa) precursor that contains a 29 aa signal peptide and a 361 aa proprotein (4). A furin-like convertase processes the proprotein within the trans-Golgi to generate an N‑terminal 249 aa (aa 30-278) latency-associated peptide (LAP) and a C-terminal 112 aa mature TGF-beta 1 (aa 279-390) (4‑6). Disulfide-linked homodimers of LAP and TGF-beta 1 remain non‑covalently associated after secretion, forming the small latent TGF-beta 1 complex (4‑8). Purified LAP is also capable of associating with active TGF-beta with high affinity, and can neutralize TGF-beta activity (9). Covalent linkage of LAP to one of three latent TGF-beta binding proteins (LTBPs) creates a large latent complex that may interact with the extracellular matrix (5‑7). TGF-beta activation from latency is controlled both spatially and temporally, by multiple pathways that include actions of proteases such as plasmin and MMP9, and/or by thrombospondin 1 or selected integrins (5, 8). The LAP portion of human TGF-beta 1 shares 91%, 92%, 85%, 86% and 88% aa identity with porcine, canine, mouse, rat and equine TGF-beta 1 LAP, respectively, while mature human TGF-beta 1 portion shares 100% aa identity with porcine, canine and bovine TGF-beta 1, and 99% aa identity with mouse, rat and equine TGF-beta 1. Although different isoforms of TGF-beta are naturally associated with their own distinct LAPs, the TGF-beta 1 LAP is capable of complexing with, and inactivating, all other human TGF-beta isoforms and those of most other species (9). Mutations within the LAP are associated with Camurati-Engelmann disease, a rare sclerosing bone dysplasia characterized by inappropriate presence of active TGF-beta 1 (10).
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