Human LAP (TGF-beta 1) Antibody

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

R&D Systems
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Key Product Details

Species Reactivity

Validated:

Human

Cited:

Human, Mouse, Rat, Porcine, Mink, Rabbit, Transgenic Mouse

Applications

Validated:

Western Blot, Neutralization, Immunocytochemistry, Simple Western

Cited:

Immunohistochemistry, Immunohistochemistry-Paraffin, Immunohistochemistry-Frozen, Western Blot, Neutralization, Immunocytochemistry, Bioassay, Dot Blot

Label

Unconjugated

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 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 direct ELISAs and Western blots.

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

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

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

Western blot shows lysates of Saos-2 human osteosarcoma cell line. PVDF membrane was probed with 2 µg/mL of Goat Anti-Human LAP (TGF-beta 1) Antigen Affinity-purified Polyclonal Antibody (Catalog # AF-246-NA) followed by HRP-conjugated Anti-Goat IgG Secondary Antibody (Catalog # HAF017). A specific band was detected for LAP (TGF-beta 1) at approximately 48 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.

LAP (TGF-beta 1) in Human PBMCs.

LAP (TGF-beta 1) was detected in immersion fixed Human peripheral blood mononuclear cells using Goat Anti-Human LAP (TGF-beta 1) Antigen Affinity-purified Polyclonal Antibody (Catalog # AF-246-NA) at 15 µg/mL for 3 hours at room temperature. Cells were stained using the NorthernLights™ 557-conjugated Anti-Goat IgG Secondary Antibody (red; NL001) and counterstained with DAPI (blue). Specific staining was localized to cytoplasm. Staining was performed using our protocol for Fluorescent ICC Staining of Non-adherent Cells.
Detection of Human LAP (TGF-beta 1) antibody by Simple WesternTM.

Detection of Human LAP (TGF-beta 1) by Simple WesternTM.

Simple Western lane view shows lysates of K562 human chronic myelogenous leukemia cell line and A549 human lung carcinoma cell line, loaded at 0.2 mg/mL. A specific band was detected for LAP (TGF-beta 1) at approximately 61 kDa (as indicated) using 20 µg/mL of Goat Anti-Human LAP (TGF-beta 1) Antigen Affinity-purified Polyclonal Antibody (Catalog # AF-246-NA) followed by 1:50 dilution of HRP-conjugated Anti-Goat IgG Secondary Antibody (Catalog # HAF109). This experiment was conducted under reducing conditions and using the 12-230 kDa separation system.
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 Recom-binant 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 Recom-binant Human TGF-beta 1 (1 ng/mL) activity elicited by Recombin-ant Human LAP TGF-beta 1 (500 ng/mL) is neutralized (green line) by increasing concentrations of Goat Anti-Human LAP TGF-beta 1 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF-246-NA). The ND50 is typically 0.4-2 µg/mL.
Detection of LAP (TGF-beta 1) by Western Blot

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

Re-expression of MYST1 inhibits TGF beta -dependent fibroblast activation and prevents experimental fibrosis.a, b Overexpression of MYST1 in cultured human dermal fibroblasts: a Expression of alpha SMA (n = 4 biological replicates per group) and formation of stress fibers (n = 5 biological replicates per group). Representative images and quantification. Horizontal scale bar, 100 µm. b mRNA levels of COL1A1 analyzed by qRT-PCR (n = 5 biological replicates per group), total collagen as analyzed by hydroxyproline assays (n = 4 biological replicates per group) and type I collagen protein as analyzed by western blot (n = 5 biological replicates per group). c–e Adenoviral overexpression of MYST1 in murine models. c T beta RIact-induced skin fibrosis: Representative trichrome stainings for, dermal thickening, myofibroblast counts and hydroxyproline content (n = 4 biological replicates per group). d Bleomycin-induced pulmonary fibrosis: Representative Trichrome stainings, fibrotic area, myofibroblast counts, Ashcroft scores and hydroxyproline content (n = 4 biological replicates for the group AdMyst1 treated with TGF beta and n = 3 for other groups). e Bleomycin-induced dermal fibrosis: Representative trichrome stainings shown, dermal thickening, myofibroblast counts and hydroxyproline content (n = 4 biological replicates for bleomycin treated groups and n = 3 for saline treated groups). Horizontal scale bars, 100 µm. All data are presented as median ± IQR. p-values were determined by ANOVA one-way with Tukey’s multiple comparison post hoc test and are indicated in the figure. See source data for more detailed information. rel.: relative, Ad: adenovirus, conc.: concentration, int.: intensity, T beta RIact: constitutively active TGF beta receptor type I, Bleo: bleomycin. Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/34285225), licensed under a CC-BY license. Not internally tested by R&D Systems.
Detection of LAP (TGF-beta 1) by Western Blot

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

Downregulation of MYST1 mediates the stimulatory effects of TGF beta on autophagy.a, b Effects of siRNA-mediated knockdown of MYST1 on autophagy in healthy human dermal fibroblasts as assessed by quantification of the mRNA levels of ATG7 (n = 4 biological replicates per group and n = 3 for MYST1 siRNA + TGF beta ), BECLIN1 (n = 4 biological replicates per group) and respective protein quantifications (n = 4 biological replicates per group). Confirmation of reduced mRNA (n = 4 biological replicates per group) (a) and protein (b; n = 4 biological replicates per group) levels of MYST1 by qRT-PCR and western blot. c EGFP/RFP-LC3 reporter activity upon knockdown of MYST1; representative images and quantification (n = 5 biological replicates per group). Horizontal scale bar, 5 µm. d, e mRNA expression of ATG7, BECLIN1, and MYST1 (n = 6 biological replicates per group) (d) and respective protein quantification (e) (n = 4 biological replicates per group) levels after adenoviral overexpression of MYST1 in healthy human dermal fibroblasts (e). All data are presented as mean ± IQR. p-values were determined by ANOVA one-way with Tukey’s multiple comparison post hoc test. See source data for more detailed information. rel.: relative, Ad: adenovirus. The results in panel c are part of a large set of experiments and share controls with Supplementary Fig. 8a. Data are presented separately just for the purpose of this figure. Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/34285225), licensed under a CC-BY license. Not internally tested by R&D Systems.

Applications for Human LAP (TGF-beta 1) Antibody

Application
Recommended Usage

Immunocytochemistry

5-15 µg/mL
Sample: Immersion fixed Human peripheral blood mononuclear cells

Simple Western

20 µg/mL
Sample: K562 human chronic myelogenous leukemia cell line and A549 human lung carcinoma cell line

Western Blot

2 µg/mL
Sample: Saos‑2 human osteosarcoma cell line

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 0.4-2 µg/mL in the presence of 500 ng/mL Recombinant Human LAP TGF‑ beta 1 and 1 ng/mL TGF‑ beta 1.

Reviewed Applications

Read 4 reviews rated 4 using AF-246-NA in the following applications:

Formulation, Preparation, and Storage

Purification

Antigen Affinity-purified

Reconstitution

Reconstitute at 0.2 mg/mL in sterile PBS. For liquid material, refer to CoA for concentration.

Formulation

Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. *Small pack size (SP) is supplied either lyophilized or as a 0.2 µm filtered solution in PBS.

Shipping

Lyophilized product is shipped at ambient temperature. Liquid small pack size (-SP) is shipped with polar packs. Upon receipt, store 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 mature TGF-beta 1 (aa 279-390) (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) Antibody

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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 (4)

4 out of 5
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  • Human LAP (TGF-beta 1) Antibody
    Name: Anonymous
    Application: Western Blot
    Sample Tested: Pancreatic cancer cells
    Species: Human
    Verified Customer | Posted 01/27/2021
    Human LAP (TGF-beta 1) Antibody AF-246-NA
  • Human LAP (TGF-beta 1) Antibody
    Name: Jeremie Raso
    Application: Western Blot
    Sample Tested: Aorta tissue
    Species: Rat
    Verified Customer | Posted 02/03/2020
    Human LAP (TGF-beta 1) Antibody AF-246-NA
  • Name: Anonymous
    Application: Western Blot
    Sample Tested: See PMID 21998117
    Species: Other
    Verified Customer | Posted 01/07/2015
  • Name: Anonymous
    Application: Western Blot
    Sample Tested: See PMID 22835852
    Species: Other
    Verified Customer | Posted 01/07/2015

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