|Detection of LAP (TGF-beta 1) in Human Platelets by Flow Cytometry. Human platelets were stained with Mouse Anti-Human LAP (TGF-beta 1) Alexa Fluor® 488‑conjugated Monoclonal Antibody (Catalog # FAB2463G, filled histogram) or isotype control antibody (Catalog # IC002G, open histogram). View our protocol for Staining Membrane-associated Proteins.|
|LAP (TGF-beta 1) in Human PBMCs. LAP (TGF-beta 1) was detected in immersion fixed human peripheral blood mononuclear cells (PBMCs) stimulated with CD3/CD28/TGF-beta/IL-2 using Mouse Anti-Human LAP (TGF-beta 1) Alexa Fluor® 488‑conjugated Monoclonal Antibody (green; Catalog # FAB2463G) at 5 µg/mL for 3 hours at room temperature. Cells were counterstained with DAPI (blue). View our protocol for Fluorescent ICC Staining of Non-adherent Cells.|
TGF-beta 1 is one member of a six gene family that has been described in mammals, birds, fish and frog. The name TGF-beta is applied to a 24-28 kDa disulfide-linked dimer that is generated through the proteolytic processing of a larger precursor molecule. For TGF-beta 1, a 50-55 kDa, 391 amino acid (aa) proprecursor is first, covalently linked to a second proprecursor (creating a disulfide-linked homodimer), and second, internally cleaved to generate two covalently-linked homodimers that remain non-covalently associated. The smallest homodimer representing aa 279-390 of the proform is TGF-beta 1; the largest homodimer representing aa 30-278 of the proform is termed LAP (Latency-associated Peptide). The LAP homodimer wraps itself around the smaller TGF-beta 1 homodimer, thus blocking an interaction of mature TGF-beta with its receptors. Almost all cells secrete the inactive TGF-beta :LAP complex, and most do so with LAP covalently bound to a very large 120-160 kDa LTBP (Latent TGF-beta Binding Protein), platelets being a notable exception. LTBP associates with multiple matrix components and this serves to store TGF-beta extracellularly in a non-active form. When TGF-beta signaling is needed, the LTBP:matrix association is disrupted, and the TGF-beta :LAP complex is exposed to multiple LAP binding partners such as TSP-1 and various Integrins. Interactions with these factors cause LAP to unwrap and dissociate from TGF-beta, resulting in TGF-beta "activation" and receptor binding. Ther are three human TGF-beta 1 LAPs and TGF-beta 1 LAP shares 36% and 33% aa sequence indentity with TGF-beta 2 LAP and TGF-beta 3 LAP, respectively. Human to mouse, TGF-beta 1 LAP shares 86% aa sequence identity.
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