Human LAP (TGF-beta 1) Quantikine ELISA Kit

Name: Human LAP (TGF-beta 1) Quantikine ELISA Kit
Brand: R&D Systems
SKU: DLAP00
Rating: 4.7 (3 reviews)

Catalog #: DLAP00
4 Citations 3 Reviews Datasheet / COA / SDS

Catalog #	Availability	Size / Price	Qty
DLAP00

Control Products Available

QC110: Quantikine Immunoassay Control Set 788 Human LAP (TGF-b1)

Ancillary Products Available

DY010: Sample Activation Kit 1

Human LAP (TGF-beta 1) ELISA Standard Curve

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All LAP (TGF-beta 1) ELISAs

Product Details

Citations (4)

FAQs

Supplemental Products

Reviews (3)

Summary Precision Recovery Linearity Data Examples Product Datasheets Preparation and Storage Background Related Research Areas

Human LAP (TGF-beta 1) Quantikine ELISA Kit Summary

Assay Length

4.5 hours

Sample Type & Volume Required Per Well

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

Sensitivity

3.4 pg/mL

Assay Range

23.4 - 1,500 pg/mL (Cell Culture Supernates, Cell Lysates, Serum, Platelet-poor EDTA Plasma, Platelet-poor Heparin Plasma, Urine)

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.

Product Summary

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).

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, Serum, Platelet-poor EDTA Plasma, Platelet-poor Heparin Plasma, 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

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.

Scientific Data

N/A

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Human LAP (TGF-beta 1) ELISA Standard Curve

Product Datasheets

Product Datasheet

COA

SDS

Preparation and Storage

Shipping

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

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 (T_reg) 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 T_reg (14, 20-23).

Long Name:

Latency-associated Peptide

Entrez Gene IDs:

7040 (Human)

Alternate Names:

CED; DPD1; LAP (TGFbeta 1); LAP (TGF-beta 1); LAP; TGFB; TGFB1; TGFbeta; transforming growth factor beta 1

Related Research Areas

⚠ 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

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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Bone marrow mesenchymal stem cells paracrine TGF-beta1 to mediate the biological activity of osteoblasts in bone repair
Authors: X Zhang, G Wang, W Wang, C Ran, F Piao, Z Ma, Z Zhang, G Zheng, F Cao, H Xie, D Cui, C Samuel Oko, X Yu, Z Wang, D Zhao
Cytokine, 2023-02-02;164(0):156139.
Species: Human
Sample Types: Cell Culture Supernates
The histone demethylase PHF8 regulates TGFbeta signaling and promotes melanoma metastasis
Authors: RS Moubarak, A de Pablos-, V Ortiz-Bara, Y Gong, M Gowen, I Dolgalev, SAA Shadaloey, D Argibay, A Karz, R Von Itter, EC Vega-Sáenz, E Sokolova, F Darvishian, A Tsirigos, I Osman, E Hernando
Science Advances, 2022-02-18;8(7):eabi7127.
Species: Human
Sample Types: Cell Culture Supernates
Characteristics of autologous protein solution and leucocyte-poor platelet-rich plasma for the treatment of osteoarthritis of the knee
Authors: S Wasai, M Sato, M Maehara, E Toyoda, R Uchiyama, T Takahashi, E Okada, Y Iwasaki, S Suzuki, M Watanabe
Sci Rep, 2020;10(1):10572.
Species: Human
Sample Types: Plasma
Effects of pirfenidone targeting the tumor microenvironment and tumor-stroma interaction as a novel treatment for non-small cell lung cancer
Authors: A Fujiwara, S Funaki, E Fukui, K Kimura, T Kanou, N Ose, M Minami, Y Shintani
Sci Rep, 2020;10(1):10900.
Species: Human
Sample Types: Cell Culture Supernates