Human LAP (TGF-beta 1) Alexa Fluor® 700-conjugated Antibody Summary
Leu30-Arg278 (Cys33Ser)
Accession # P01137
Applications
Please Note: Optimal dilutions should be determined by each laboratory for each application. General Protocols are available in the Technical Information section on our website.
Scientific Data
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® 700-conjugated Monoclonal Antibody (Catalog # FAB2463N, filled histogram) or isotype control antibody (Catalog # IC002N, open histogram). Histograms were gated on CD41+cells. View our protocol for Staining Membrane-associated Proteins.
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Preparation and Storage
Background: LAP (TGF-beta 1)
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.
Product Datasheets
Product Specific Notices
This product is provided under an agreement between Life Technologies Corporation and R&D Systems, Inc, and the manufacture, use, sale or import of this product is subject to one or more US patents and corresponding non-US equivalents, owned by Life Technologies Corporation and its affiliates. The purchase of this product conveys to the buyer the non-transferable right to use the purchased amount of the product and components of the product only in research conducted by the buyer (whether the buyer is an academic or for-profit entity). The sale of this product is expressly conditioned on the buyer not using the product or its components (1) in manufacturing; (2) to provide a service, information, or data to an unaffiliated third party for payment; (3) for therapeutic, diagnostic or prophylactic purposes; (4) to resell, sell, or otherwise transfer this product or its components to any third party, or for any other commercial purpose. Life Technologies Corporation will not assert a claim against the buyer of the infringement of the above patents based on the manufacture, use or sale of a commercial product developed in research by the buyer in which this product or its components was employed, provided that neither this product nor any of its components was used in the manufacture of such product. For information on purchasing a license to this product for purposes other than research, contact Life Technologies Corporation, Cell Analysis Business Unit, Business Development, 29851 Willow Creek Road, Eugene, OR 97402, Tel: (541) 465-8300. Fax: (541) 335-0354.
FAQs
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How do you isolate human platelets for flow cytometry, using Human LAP (TGF-beta 1) Antibody, Catalog # MAB2463, and its conjugates?
We obtain the platelets from blood plasma by collection the top layer after spinning down whole blood. We proceed with a normal surface stain protocol; no special buffers are required.
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