Human LAP TGF-beta 1 Biotinylated Antibody Summary
Leu30-Ser390
Accession # P01137
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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
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Detection of Human LAP (TGF-beta 1) by Western Blot Disulfide-linked GARP/TGF-beta 1 complexes are released in the supernatant of T cells, but not 293 cells.A. Cells described in Figure 2 were lysed and immunoprecipitated (IP) with anti-GARP or anti-LAP antibodies. IP products were submitted to SDS-PAGE under non-reducing or reducing conditions, followed by WB with anti-LAP antibodies (top and middle panels), or anti-GARP antibodies (bottom panels). Pro-TGF-beta 1 and LAP homodimers in the top panels are not clearly resolved, but can be distinguished better with longer migrations or higher concentrations of polyacrylamide. The +/- 85-90 kDa bands that appear in the middle panel correspond to non-specific bands, or to incompletely reduced pro-TGF-beta 1. B. Cells (2x106/ml for murine and human T cells, 2.5x105/ml for transfected 293 cells) were incubated in serum free medium during 24 hours. Different cell concentrations were used to adjust for the different amounts of secreted TGF-beta 1 (see Figure 2). Human Th A2 and Jurkat cells were stimulated with anti-CD3/CD28 antibodies to increase secretion. Supernatants (0.5-10 µl) were analyzed by WB under non-reducing conditions with anti-GARP and anti-LAP antibodies. * Band that also appears when the secondary anti-IgG2b-HRP antibody is used alone (without anti-GARP antibody), due to cross reactivity against the anti-CD3/CD28 antibodies used for T cell stimulation. Image collected and cropped by CiteAb from the following publication (https://dx.plos.org/10.1371/journal.pone.0076186), licensed under a CC-BY license. Not internally tested by R&D Systems.
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Detection of Human LAP (TGF-beta 1) by Western Blot Disulfide-linked GARP/TGF-beta 1 complexes are released by stimulated human Tregs, which naturally express GARP.The indicated Treg and Th cell populations were left resting or stimulated with anti-CD3/CD28 antibodies in serum-free medium. A. Supernatants were collected after 48 hours. B. Cell lysates were collected after 24 hours and IP with an anti-GARP antibody. Supernatants (A) and immunoprecipitated lysates (B) were submitted to SDS-PAGE under non-reducing conditions, then analyzed by WB with anti-LAP antibodies. Similar results were obtained with freshly isolated CD4+ T cells from 2 other donors and with expanded CD4+ T cells from 5 others donors. Image collected and cropped by CiteAb from the following publication (https://dx.plos.org/10.1371/journal.pone.0076186), licensed under a CC-BY license. Not internally tested by R&D Systems.
Preparation and Storage
- 12 months from date of receipt, -20 to -70 °C as supplied.
- 1 month, 2 to 8 °C under sterile conditions after reconstitution.
- 6 months, -20 to -70 °C under sterile conditions after reconstitution.
Background: LAP (TGF-beta 1)
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 (aa 279-390) mature TGF- beta 1 (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).
- Dunker, N. & K. Krieglstein (2000) Eur. J. Biochem. 267:6982.
- Wahl, S.M. (2006) Immunol. Rev. 213:213.
- Chang, H. et al. (2002) Endocr. Rev. 23:787.
- Derynck, R. et al. (1985) Nature 316:701.
- Dabovic, B. and D.B. Rifkin (2008) “TGF-beta Bioavailability” in The TGF-beta Family. Derynck, R. and K. Miyazono (eds): Cold Spring Harbor Laboratory Press, p. 179.
- Brunner, A.M. et al. (1989) J. Biol. Chem. 264:13660.
- Miyazono, K. et al. (1991) EMBO J. 10:1091.
- Oklu, R. and R. Hesketh (2000) Biochem. J. 352:601.
- Miller, D.M. et al. (1992) Mol. Endocrinol. 6:694.
- Janssens, K. et al. (2003) J. Biol. Chem. 278:7718.
Product Datasheets
Citations for Human LAP TGF-beta 1 Biotinylated Antibody
R&D Systems personnel manually curate a database that contains references using R&D Systems products. The data collected includes not only links to publications in PubMed, but also provides information about sample types, species, and experimental conditions.
17
Citations: Showing 1 - 10
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LRRC33 is a novel binding and potential regulating protein of TGF-beta 1 function in human acute myeloid leukemia cells
Authors: Wenjiang Ma, Yan Qin, Bjoern Chapuy, Chafen Lu
PLOS ONE
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A Milieu Molecule for TGF-beta Required for Microglia Function in the Nervous System
Authors: Yan Qin, Brian S. Garrison, Wenjiang Ma, Rui Wang, Aiping Jiang, Jing Li et al.
Cell
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Hsp65-Producing Lactococcus lactis Prevents Inflammatory Intestinal Disease in Mice by IL-10- and TLR2-Dependent Pathways
Authors: Ana Cristina Gomes-Santos, Rafael Pires de de Oliveira, Thaís Garcias Moreira, Archimedes Barbosa Castro-Junior, Bernardo Coelho Horta, Luísa Lemos et al.
Frontiers in Immunology
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Lysosomal-associated Transmembrane Protein 4B (LAPTM4B) Decreases Transforming Growth Factor beta1 (TGF-beta1) Production in Human Regulatory T Cells.
Authors: Huygens C, Lienart S, Dedobbeleer O, Stockis J, Gauthy E, Coulie P, Lucas S
J Biol Chem, 2015-06-30;290(33):20105-16.
Species: Human
Sample Types: Cell Lysates
Applications: Western Blot -
GARP Is Regulated by miRNAs and Controls Latent TGF-beta 1 Production by Human Regulatory T Cells
Authors: Emilie Gauthy, Julia Cuende, Julie Stockis, Caroline Huygens, Bernard Lethé, Jean-François Collet et al.
PLoS ONE
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GARP regulates the bioavailability and activation of TGF beta
Authors: Rui Wang, Jianghai Zhu, Xianchi Dong, Minlong Shi, Chafen Lu, Timothy A. Springer
Molecular Biology of the Cell
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Inducible CD4+LAP+ Foxp3 negative Regulatory T cells Suppress Allergic Inflammation
Authors: Wei Duan, Takanori So, Amit K. Mehta, Heonsik Choi, Michael Croft
The Journal of Immunology
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Antigen-specific prevention of type 1 diabetes in NOD mice is ameliorated by OX40 agonist treatment
Authors: Damien Bresson, Georgia Fousteri, Yulia Manenkova, Michael Croft, Matthias von Herrath
Journal of Autoimmunity
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TGF-beta Induces Surface LAP Expression on Murine CD4 T Cells Independent of Foxp3 Induction
Authors: Takatoku Oida, Howard L. Weiner
PLoS ONE
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Overexpression of TGF-ß 1 gene induces cell surface localized glucose-regulated protein 78-associated latency-associated peptide/TGF-ß.
Authors: Oida T, Weiner HL
J. Immunol., 2010-08-18;185(6):3529-35.
Species: Human
Sample Types: Cell Lysates
Applications: Western Blot -
Importance of TLR2 in the direct response of T lymphocytes to Schistosoma mansoni antigens.
Authors: Burton OT, Gibbs S, Miller N, Jones FM, Wen L, Dunne DW, Cooke A, Zaccone P
Eur. J. Immunol., 2010-08-01;40(8):2221-9.
Species: Mouse
Sample Types: Whole Cells
Applications: Flow Cytometry -
Involvement of regulatory T cells in the immunosuppression characteristic of patients with paracoccidioidomycosis.
Authors: Ferreira MC, de Oliveira RT, da Silva RM, Blotta MH, Mamoni RL
Infect. Immun., 2010-07-19;78(10):4392-401.
Species: Human
Sample Types: Whole Cells
Applications: Flow Cytometry -
Stromal regulation of vessel stability by MMP14 and TGFbeta
Authors: Nor E. Sounni, Kerstin Dehne, Leon van Kempen, Mikala Egeblad, Nesrine I. Affara, Ileana Cuevas et al.
Disease Models & Mechanisms
Species: Mouse
Sample Types: Tissue Homogenates
Applications: Immunoprecipitation -
Membrane protein GARP is a receptor for latent TGF-beta on the surface of activated human Treg.
Authors: Stockis J, Colau D, Coulie PG, Lucas S
Eur. J. Immunol., 2009-12-01;39(12):3315-22.
Species: Human
Sample Types: Cell Lysates, Whole Cells
Applications: Flow Cytometry, Western Blot -
T-cell-expressed proprotein convertase furin is essential for maintenance of peripheral immune tolerance.
Authors: Pesu M, Watford WT, Wei L, Xu L, Fuss I, Strober W, Andersson J, Shevach EM, Quezado M, Bouladoux N, Roebroek A, Belkaid Y, Creemers J, O'Shea JJ
Nature, 2008-09-11;455(7210):246-50.
Species: Mouse
Sample Types: Whole Cells
Applications: Flow Cytometry -
Ig-reactive CD4+CD25+ T cells from tolerized (New Zealand Black x New Zealand White)F1 mice suppress in vitro production of antibodies to DNA.
Authors: La Cava A, Ebling FM, Hahn BH
J. Immunol., 2004-09-01;173(5):3542-8.
Species: Mouse
Sample Types: Whole Cells
Applications: Flow Cytometry -
Metastable tolerance to rhesus monkey renal transplants is correlated with allograft TGF-beta 1+CD4+ T regulatory cell infiltrates.
Authors: Torrealba JR, Katayama M, Fechner JH, Jankowska-Gan E, Kusaka S, Schultz JM, Hu H, Hamawy MM, Jonker M, Wubben J, Doxiadis G, Bontrop R, Burlingham WJ, Knechtle SJ
J. Immunol., 2004-05-01;172(9):5753-64.
Species: Primate - Macaca mulatta (Rhesus Macaque)
Sample Types: Tissue Homogenates, Whole Tissue
Applications: IHC-P, Western Blot
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