LAP (TGF-beta 1) Antibody [Biotin]

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 (//dx.plos.org/10.1371/journal.pone.0076186), licensed under a CC-BY license. Not internally tested by R&D Systems.

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 (//dx.plos.org/10.1371/journal.pone.0076186), licensed under a CC-BY license. Not internally tested by R&D Systems.

• Reactivity: Hu
• Applications: WB
• Clonality: Polyclonal
• Host: Goat
• Conjugate: Biotin
• Concentration: LYOPH

LAP (TGF-beta 1) Antibody [Biotin] Summary

• Immunogen: S. frugiperda insect ovarian cell line Sf 21-derived recombinant human LAP TGF‑ beta 1 and Chinese hamster ovary cell line CHO-derived recombinant human LAP TGF‑ beta 1
  Leu30-Ser390
  Accession # P01137
• Specificity: Detects human LAP TGF-beta 1 in Western blots. In this format, less than 1% cross-reactivity with mature recombinant human (rh) TGF‑ beta 1, porcine TGF‑ beta 2, rhTGF‑ beta 3, and recombinant amphibian TGF‑ beta 5 is observed.
• Source: N/A
• Isotype: IgG
• Clonality: Polyclonal
• Host: Goat
• Gene: TGFB1
• Purity Statement: Antigen Affinity-purified

Applications/Dilutions

• Dilutions:
  • Western Blot 0.1 ug/mL
• Readout System:
  • Streptavidin Native Protein NBP2-29367
  • Streptavidin Native Protein NBP2-29370
  • Streptavidin Native Protein NBP2-29371

Packaging, Storage & Formulations

• Storage: Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
  • 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.
• Buffer: Lyophilized from a 0.2 μm filtered solution in PBS with BSA as a carrier protein.
• Preservative: No Preservative
• Concentration: LYOPH
• Reconstitution Instructions: Reconstitute at 0.2 mg/mL in sterile PBS.

Notes

This product is produced by and ships from R&D Systems, Inc., a Bio-Techne brand.

Alternate Names for LAP (TGF-beta 1) Antibody [Biotin]

• CED
• DPD1
• LAP (TGFbeta 1)
• LAP (TGF-beta 1)
• LAP
• TGFB
• TGFB1
• TGFbeta
• transforming growth factor beta 1

Background

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).

1. Dunker, N. & K. Krieglstein (2000) Eur. J. Biochem. 267:6982.
2. Wahl, S.M. (2006) Immunol. Rev. 213:213.
3. Chang, H. et al. (2002) Endocr. Rev. 23:787.
4. Derynck, R. et al. (1985) Nature 316:701.
5. 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.
6. Brunner, A.M. et al. (1989) J. Biol. Chem. 264:13660.
7. Miyazono, K. et al. (1991) EMBO J. 10:1091.
8. Oklu, R. and R. Hesketh (2000) Biochem. J. 352:601.
9. Miller, D.M. et al. (1992) Mol. Endocrinol. 6:694.
10. Janssens, K. et al. (2003) J. Biol. Chem. 278:7718.

Limitations

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Primary Antibodies are guaranteed for 1 year from date of receipt.

Publications for LAP (TGF-beta 1) Antibody (BAF246)(17)

Publications using BAF246

• Wenjiang Ma, Yan Qin, Bjoern Chapuy, Chafen Lu LRRC33 is a novel binding and potential regulating protein of TGF-beta 1 function in human acute myeloid leukemia cells PLOS ONE 2019-10-10 [PMID: 31600200]
• Yan Qin, Brian S. Garrison, Wenjiang Ma, Rui Wang, Aiping Jiang, Jing Li et al. A Milieu Molecule for TGF-beta Required for Microglia Function in the Nervous System Cell 2018-06-14 [PMID: 29909984]
• Ana Cristina Gomes-Santos, Rafael Pires de de Oliveira, Thaís Garcias Moreira, Archimedes Barbosa Castro-Junior, Bernardo Coelho Horta, Luísa Lemos et al. Hsp65-Producing Lactococcus lactis Prevents Inflammatory Intestinal Disease in Mice by IL-10- and TLR2-Dependent Pathways Frontiers in Immunology 2017-01-30 [PMID: 28194152]
• Huygens C, Lienart S, Dedobbeleer O, Stockis J, Gauthy E, Coulie P, Lucas S Lysosomal-associated Transmembrane Protein 4B (LAPTM4B) Decreases Transforming Growth Factor beta1 (TGF-beta1) Production in Human Regulatory T Cells. J Biol Chem, 2015-06-30;290(33):20105-16. 2015-06-30 [PMID: 26126825] (Western Blot, Human)
• Emilie Gauthy, Julia Cuende, Julie Stockis, Caroline Huygens, Bernard Lethé, Jean-François Collet et al. GARP Is Regulated by miRNAs and Controls Latent TGF-beta 1 Production by Human Regulatory T Cells PLoS ONE 2013-09-30 [PMID: 24098777]
• Rui Wang, Jianghai Zhu, Xianchi Dong, Minlong Shi, Chafen Lu, Timothy A. Springer GARP regulates the bioavailability and activation of TGF beta Molecular Biology of the Cell 2012-03-15 [PMID: 22278742]
• Wei Duan, Takanori So, Amit K. Mehta, Heonsik Choi, Michael Croft Inducible CD4+LAP+ Foxp3 negative Regulatory T cells Suppress Allergic Inflammation The Journal of Immunology 2011-12-15 [PMID: 22079987]
• Damien Bresson, Georgia Fousteri, Yulia Manenkova, Michael Croft, Matthias von Herrath Antigen-specific prevention of type 1 diabetes in NOD mice is ameliorated by OX40 agonist treatment Journal of Autoimmunity 2011-12-01 [PMID: 22063316]
• Takatoku Oida, Howard L. Weiner TGF-beta Induces Surface LAP Expression on Murine CD4 T Cells Independent of Foxp3 Induction PLoS ONE 2010-11-24 [PMID: 21124798]
• Oida T, Weiner HL Overexpression of TGF-ß 1 gene induces cell surface localized glucose-regulated protein 78-associated latency-associated peptide/TGF-ß. J. Immunol., 2010-08-18;185(6):3529-35. 2010-08-18 [PMID: 20720212] (Western Blot, Human)
• Ferreira MC, de Oliveira RT, da Silva RM, Blotta MH, Mamoni RL Involvement of regulatory T cells in the immunosuppression characteristic of patients with paracoccidioidomycosis. Infect. Immun., 2010-07-19;78(10):4392-401. 2010-07-19 [PMID: 20643858] (Flow Cytometry, Human)
• Burton OT, Gibbs S, Miller N, Jones FM, Wen L, Dunne DW, Cooke A, Zaccone P Importance of TLR2 in the direct response of T lymphocytes to Schistosoma mansoni antigens. Eur. J. Immunol., 2010-08-01;40(8):2221-9. 2010-08-01 [PMID: 20480503] (Flow Cytometry, Mouse)
• Nor E. Sounni, Kerstin Dehne, Leon van Kempen, Mikala Egeblad, Nesrine I. Affara, Ileana Cuevas et al. Stromal regulation of vessel stability by MMP14 and TGFbeta Disease Models & Mechanisms 2010-04-28 [PMID: 20223936] (Immunoprecipitation, Mouse)
• Stockis J, Colau D, Coulie PG, Lucas S Membrane protein GARP is a receptor for latent TGF-beta on the surface of activated human Treg. Eur. J. Immunol., 2009-12-01;39(12):3315-22. 2009-12-01 [PMID: 19750484] (Flow Cytometry, Western Blot, Human)
• 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 T-cell-expressed proprotein convertase furin is essential for maintenance of peripheral immune tolerance. Nature, 2008-09-11;455(7210):246-50. 2008-09-11 [PMID: 18701887] (Flow Cytometry, Mouse)
• La Cava A, Ebling FM, Hahn BH 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. J. Immunol., 2004-09-01;173(5):3542-8. 2004-09-01 [PMID: 15322219] (Flow Cytometry, Mouse)
• 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 Metastable tolerance to rhesus monkey renal transplants is correlated with allograft TGF-beta 1+CD4+ T regulatory cell infiltrates. J. Immunol., 2004-05-01;172(9):5753-64. 2004-05-01 [PMID: 15100322] (IHC-P, Western Blot, Primate - Macaca mulatta (Rhesus Macaque))

Bioinformatics

• Gene Symbol: TGFB1
• Uniprot:
  • P01137(Human)