Human LAP (TGF-beta 1) Antibody

(4 citations)   
  • Species Reactivity
    Human
  • Specificity
    Detects human LAP in direct ELISAs and Western blots. In direct ELISAs and Western blots, this antibody is specific for the LAP derived from the TGF-beta 1 precursor but does not react with LAP from the TGF-beta 2 precursor.
  • Source
    Polyclonal Goat IgG
  • Purification
    Protein A or G purified
  • 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
  • Formulation
    Lyophilized from a 0.2 μm filtered solution in PBS and NaCl with Trehalose.
  • Endotoxin Level
    <0.10 EU per 1 μg of the antibody by the LAL method.
  • Label
    Unconjugated
Applications
  •  
    Recommended
    Concentration
    Sample
  • Western Blot
    1 µg/mL
    Recombinant Human LAP TGF-beta 1 (Catalog # 246-LP)
  • Immunohistochemistry
    5-15 µg/mL
    See below
  • Immunocytochemistry
    5-15 µg/mL
    See below
  • Neutralization
    Measured by its ability to neutralize LAP TGF‑ beta 1 inhibition of TGF‑ beta 1 growth inhibition in the HT‑2 mouse T cell line. Tsang, M. et al. (1995) Cytokine 7:389. The Neutralization Dose (ND50) is typically 15-30 µg/mL in the presence of 125 ng/mL Recombinant Human LAP TGF‑ beta 1 and 0.25 ng/mL TGF‑ beta 1.
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.
Data Examples
Immunocytochemistry
TGF-beta 1 in Human PBMCs. TGF-beta 1 was detected in immersion fixed human peripheral blood mononuclear cells (PBMCs) using Goat Anti-Human LAP TGF-beta 1 Polyclonal Antibody (Catalog # AB-246-NA) at 10 µg/mL for 3 hours at room temperature. Cells were stained using the NorthernLights™ 557-conjugated Anti-Goat IgG Secondary Antibody (yellow; Catalog # NL001) and counter­stained with DAPI (blue).View our protocol for Fluorescent ICC Staining of Non-adherent Cells.
Immunohistochemistry
LAP (TGF-beta 1) in Human Prostate Cancer Tissue. LAP (TGF-beta 1) was detected in immersion fixed paraffin-embedded sections of human prostate cancer tissue using Goat Anti-Human LAP (TGF-beta 1) Antigen Affinity-purified Polyclonal Antibody (Catalog # AF-246-NA) at 10 µg/mL overnight at 4 °C. Tissue was stained using the Anti-Goat HRP-DAB Cell & Tissue Staining Kit (brown; Catalog # CTS008) and counterstained with hematoxylin (blue). Specific staining was localized to cytoplasm in epithelial cells. View our protocol for Chromogenic IHC Staining of Paraffin-embedded Tissue Sections.
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 (0.25 ng/mL) activity elicited by Recombinant Human LAP TGF‑ beta 1 (125 ng/mL) is neutral­ized (green line) by increasing concentrations of Goat Anti-Human LAP TGF‑ beta 1 Polyclonal Antibody (Catalog # AB-246-NA). The ND50 is typically 15‑30 µg/mL.
Preparation and Storage
  • Reconstitution
    Reconstitute at 1 mg/mL in sterile PBS.
  • Shipping
    The product is shipped at ambient temperature. Upon receipt, store it immediately at the temperature recommended below.
  • Stability & 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.
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).

  • References:
    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.
  • Long Name:
    Latency-associated Peptide
  • Entrez Gene IDs:
    7040 (Human)
  • Alternate Names:
    LAP (TGFbeta 1); LAP (TGF-beta 1)
Related Research Areas
Citations:

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.

4 Citations: Showing 1 - 4
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Species
Applications
Sample Type
  1. Identification of genetic loci that control mammary tumor susceptibility through the host microenvironment.
    Authors: Zhang P, Lo A, Huang Y, Huang G, Liang G, Mott J, Karpen G, Blakely E, Bissell M, Barcellos-Hoff M, Snijders A, Mao J
    Sci Rep, 2015;5(0):8919.
    Species: Mouse
    Sample Type: Whole Tissue
    Application: IHC
  2. Knockdown of prolyl-4-hydroxylase domain 2 inhibits tumor growth of human breast cancer MDA-MB-231 cells by affecting TGF-beta1 processing.
    Authors: Wottawa M, Leisering P, Ahlen M, Schnelle M, Vogel S, Malz C, Bordoli M, Camenisch G, Hesse A, Napp J, Alves F, Kristiansen G, Farhat K, Katschinski D
    Int J Cancer, 2014;132(12):2787-98.
    Species: Human
    Sample Type: Cell Lysates
    Application: WB
  3. Transglutaminase 2 cross-linking activity is linked to invadopodia formation and cartilage breakdown in arthritis.
    Authors: Lauzier A, Charbonneau M, Paquette M, Harper K, Dubois CM
    Arthritis Res. Ther., 2012;14(4):R159.
    Species: Human
    Sample Type: Cell Lysates
    Application: WB
  4. Tumor metastasis in an orthotopic murine model of head and neck cancer: possible role of TGF-beta 1 secreted by the tumor cells.
    Authors: Dasgupta S, Bhattacharya-Chatterjee M, O&amp;amp;apos;Malley BW, Chatterjee SK
    J. Cell. Biochem., 2006;97(5):1036-51.
    Species: Mouse
    Sample Type: Whole Tissue
    Application: IHC Frozen
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