Key Product Details

Species Reactivity

Validated:

Human

Cited:

Human, Mouse, Primate - Macaca mulatta (Rhesus Macaque)

Applications

Validated:

Western Blot

Cited:

Immunohistochemistry-Paraffin, Western Blot, Flow Cytometry, Immunoprecipitation

Label

Biotin

Antibody Source

Polyclonal Goat IgG
View all LAP (TGF-beta 1) Primary Antibodies »

Product Specifications

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.

Clonality

Polyclonal

Host

Goat

Isotype

IgG

Scientific Data Images for Human LAP TGF‑ beta 1 Biotinylated Antibody

Detection of Human LAP (TGF-beta 1) by Western Blot

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.
Detection of Human LAP (TGF-beta 1) by Western Blot

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.

Applications for Human LAP TGF‑ beta 1 Biotinylated Antibody

Application
Recommended Usage

Western Blot

0.1 µg/mL
Sample: Recombinant Human LAP TGF-beta 1 (Catalog # 246-LP)

Formulation, Preparation, and Storage

Purification

Antigen Affinity-purified

Reconstitution

Reconstitute at 0.2 mg/mL in sterile PBS.

Formulation

Lyophilized from a 0.2 μm filtered solution in PBS with BSA as a carrier protein.

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.

Calculators

The reconstitution calculator allows you to quickly calculate the volume of a reagent to reconstitute your vial. Simply enter the mass of reagent and the target concentration and the calculator will determine the rest.

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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

Alternate Names

LAP (TGFbeta 1)

Entrez Gene IDs

7040 (Human)

Gene Symbol

TGFB1

UniProt

P01137 (Human)

Additional LAP (TGF-beta 1) Products

Product Documents for Human LAP TGF‑ beta 1 Biotinylated Antibody

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Product Specific Notices for Human LAP TGF‑ beta 1 Biotinylated Antibody

For research use only

Citations for Human LAP TGF‑ beta 1 Biotinylated Antibody

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