Human LAP (TGF-beta 1) Antibody

R&D Systems | Catalog # AB-246-NA

R&D Systems
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Key Product Details

Species Reactivity

Validated:

Human

Cited:

Human, Mouse

Applications

Validated:

Immunohistochemistry, Western Blot, Neutralization, Immunocytochemistry

Cited:

Immunohistochemistry, Immunohistochemistry-Paraffin, Immunohistochemistry-Frozen, Western Blot, Neutralization

Label

Unconjugated

Antibody Source

Polyclonal Goat IgG
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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 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.

Clonality

Polyclonal

Host

Goat

Isotype

IgG

Endotoxin Level

<0.10 EU per 1 μg of the antibody by the LAL method.

Scientific Data Images for Human LAP (TGF-beta 1) Antibody

TGF-beta 1 antibody in Human PBMCs by Immunocytochemistry (ICC).

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 counter-stained with DAPI (blue).View our protocol for Fluorescent ICC Staining of Non-adherent Cells.

LAP TGF‑ beta 1 Inhibition of TGF‑ beta 1 Activity and Neutralization by Human LAP TGF‑ beta 1 Antibody.

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 neutral-ized (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.

LAP (TGF-beta 1) antibody in Human Prostate Cancer Tissue by Immunohistochemistry (IHC-P).

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.

Applications for Human LAP (TGF-beta 1) Antibody

Application
Recommended Usage

Immunocytochemistry

5-15 µg/mL
Sample: Immersion fixed human peripheral blood mononuclear cells (PBMCs)

Immunohistochemistry

5-15 µg/mL
Sample: Immersion fixed paraffin-embedded sections of human prostate cancer tissue

Western Blot

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

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.

Reviewed Applications

Read 1 review rated 5 using AB-246-NA in the following applications:

Formulation, Preparation, and Storage

Purification

Protein A or G purified

Reconstitution

Reconstitute at 1 mg/mL in sterile PBS.


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Formulation

Lyophilized from a 0.2 μm filtered solution in PBS and NaCl with Trehalose.

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

Additional LAP (TGF-beta 1) Products

Product Documents for Human LAP (TGF-beta 1) Antibody

Certificate of Analysis

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Note: Certificate of Analysis not available for kit components.

Product Specific Notices for Human LAP (TGF-beta 1) Antibody

For research use only

Citations for Human LAP (TGF-beta 1) Antibody

Customer Reviews for Human LAP (TGF-beta 1) Antibody (1)

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  • Name: Anonymous
    Application: Western Blot
    Sample Tested: See PMID 22334695
    Species: Human
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