Human LAP (TGF-beta 1) Quantikine ELISA Kit
R&D Systems, part of Bio-Techne | Catalog # DLAP00
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)
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
Reactivity
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.
Scientific Data Images for Human LAP (TGF-beta 1) Quantikine ELISA Kit
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)
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
Product Specific Notices for Human LAP (TGF-beta 1) Quantikine ELISA Kit
For research use only
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⚠ 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.