Human LAP (TGF-beta 1) Quantikine ELISA Kit

R&D Systems | Catalog # DLAP00

R&D Systems
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Key Product Details

Assay Length

4.5 hours

Sample Type & Volume Required Per Well

Cell Culture Supernates (50 µL), Cell Lysates (50 µL), Serum (10 µL), Platelet-poor EDTA Plasma (10 µL), Platelet-poor Heparin Plasma (10 µL), Urine (50 µL)

Sensitivity

3.4 pg/mL

Assay Range

23.4-1500 pg/mL (Cell Culture Supernates, Cell Lysates, Serum, Platelet-poor EDTA Plasma, Platelet-poor Heparin Plasma, Urine)
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Product Summary for Human LAP (TGF-beta 1) Quantikine ELISA Kit

The Quantikine Human LAP (TGF-beta 1) Immunoassay is a 4.5 hour solid phase ELISA designed to measure LAP (TGF-beta 1) in acid activated cell culture supernates, serum, plasma, and urine. It contains recombinant human LAP (TGF-beta 1) expressed by CHO cells and has been shown to quantitate the recombinant factor accurately. Results obtained using natural LAP (TGF-beta 1) showed linear curves that were parallel to the standard curves obtained using the recombinant kit standards. These results indicate that this kit will provide accurate quantitation for both recombinant and natural human LAP (TGF-beta 1).

Product Specifications

Measurement

Quantitative ELISA

Detection Method

Colorimetric - 450nm (TMB)

Conjugate

HRP

Species

Human

Specificity

Natural and recombinant human LAP (TGF-beta 1) and Latent TGF-beta 1).

Cross-reactivity

< 0.5% cross-reactivity observed with available related molecules. < 50% cross-species reactivity observed with species tested.

Interference

No significant interference observed with available related molecules.

Precision

Intra-Assay Precision (Precision within an assay) Three samples of known concentration were tested on one plate to assess intra-assay precision.

Inter-Assay Precision (Precision between assays) Three samples of known concentration were tested in separate assays to assess inter-assay precision. Assays were performed by at least three technicians using two lots of components.

Cell Culture Supernates, Cell Lysates, Platelet-poor EDTA Plasma, Platelet-poor Heparin Plasma, Serum, Urine

Intra-Assay Precision Inter-Assay Precision
Sample 1 2 3 1 2 3
n 20 20 20 20 20 20
Mean (pg/mL) 218 427 992 216 460 1126
Standard Deviation 6.92 17.7 42.1 7.68 21.7 86.9
CV% 3.2 4.1 4.2 3.6 4.7 7.7

Recovery for Human LAP (TGF-beta 1) Quantikine ELISA Kit

The recovery of LAP (TGF-beta 1) spiked to levels throughout the range of the assay followed by activation was evaluated.

Sample Type Average % Recovery Range %
Cell Culture Supernates (n=4) 98 90-105
Cell Lysates (n=4) 97 90-103
Urine (n=4) 95 87-101

Linearity

To assess the linearity of the assay, activated samples containing and/or spiked with high concentrations of LAP (TGF-beta 1) were diluted with Calibrator Diluent and then assayed.

Human LAP (TGF-beta 1) ELISA Linearity

Scientific Data Images for Human LAP (TGF-beta 1) Quantikine ELISA Kit

Human LAP (TGF-beta 1) ELISA Standard Curve

Human LAP (TGF-beta 1) ELISA Standard Curve

Preparation and Storage

Shipping

The product is shipped at ambient temperature. Upon receipt, store it immediately at the temperature recommended below.

Stability & Storage

Store the unopened product at 2 - 8 °C. Do not use past expiration date.

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 with distinct, but overlapping, activities regulating processes such as immune function, proliferation, and epithelial mesenchymal transition (1-3). Almost all cells can produce TGF-beta 1, and a particularly high concentration is found in platelets (1-4). Human TGF-beta 1 cDNA encodes a 390 amino acid (aa) precursor that contains a 29 aa signal peptide and a 361 aa proprotein (2). A furin-like convertase processes the proprotein within the trans-Golgi to generate an N-terminal 249 aa latency-associated peptide (LAP) and a C-terminal 112 aa mature TGF- beta 1 (2, 3, 5). Disulfide-linked homodimers of LAP and TGF-beta 1 remain non-covalently associated after secretion, forming the inactive small latent TGF-beta 1 complex (5-12). 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 (3, 5-7, 13). TGF-beta activation from latency is controlled both spatially and temporally by multiple pathways that may include the actions of proteases such as plasmin, MMP-2 or MMP-9, adhesion through integrins such as alpha v beta 6 and alpha v beta 8, and/or interaction with Thrombospondin-1 (3, 5, 7, 12-14). 
Recombinant TGF-beta 1 LAP is capable of complexing with and inactivating all other human TGF-beta isoforms and those of most other species (15). The TGF-beta 1 LAP is associated in vivo with TGF-beta 1 and binds >98% of the TGF-beta 1 in circulation (16). Its circulating concentration correlates with that of total TGF-beta 1, making LAP a convenient surrogate in the measurement of TGF-beta 1 (16). LAP is accessible for antibody recognition whether or not TGF-beta 1 is bound. In contrast, latent TGF-beta 1 is surrounded by a LAP "straightjacket", altering its conformation and sequestering it from recognition by receptors or antibodies (4, 8). Acid dissociation of TGF-beta 1 from its LAP is thus required to assay total TGF-beta 1 (4, 16). 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 the 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. Assays for TGF-beta 1 LAP may therefore be more species-specific than assays for mature TGF-beta 1. Assay of human LAP is an especially convenient way to eliminate interference from bovine serum TGF-beta 1 and its LAP in tissue culture media. 
Deletion of the mouse TGF-beta 1 gene is lethal, either due to defective prenatal hematopoiesis and endothelial differentiation, or to postnatal overwhelming inflammation (17). Mutations within the human TGF-beta 1 LAP are associated with Camurati-Engelmann disease, a rare sclerosing bone dysplasia characterized by inappropriate presence of active TGF-beta 1 (7-9, 18). LAP on the surface of immature dendritic cells promotes differentiation and survival of regulatory T cells (Treg) while inhibiting Th1 cell differentiation (19). Latent TGF-beta 1 is also expressed on the surface of Treg, where it is immobilized by binding of LAP to the alternate latent TGF binding protein, GARP, and contributes to the immunosuppressive role of Treg (14, 20-23).

Long Name

Latency-associated Peptide

Alternate Names

LAP (TGFbeta 1)

Entrez Gene IDs

7040 (Human)

Gene Symbol

TGFB1

Additional LAP (TGF-beta 1) Products

Product Documents for Human LAP (TGF-beta 1) Quantikine ELISA Kit

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) Quantikine ELISA Kit

For research use only

⚠ WARNING: This product can expose you to chemicals including N,N-Dimethylforamide, which is known to the State of California to cause cancer. For more information, go to www.P65Warnings.ca.gov.

Citations for Human LAP (TGF-beta 1) Quantikine ELISA Kit

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  • Human LAP (TGF-beta 1) Quantikine ELISA Kit
    Name: Anonymous
    Sample Tested: Peripheral blood mononuclear cells (PBMCs)
    Verified Customer | Posted 01/16/2020
    Human LAP (TGF-beta 1) Quantikine ELISA Kit DLAP00
  • Human LAP (TGF-beta 1) Quantikine ELISA Kit
    Name: Anonymous
    Sample Tested: Purified Standard
    Verified Customer | Posted 09/20/2019
    Human LAP (TGF-beta 1) Quantikine ELISA Kit DLAP00
  • Human LAP (TGF-beta 1) Quantikine ELISA Kit
    Name: Anonymous
    Sample Tested: Blood mononuclear cells (PBMCs)
    Verified Customer | Posted 03/21/2019

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