Human/Mouse/Rat/Porcine/Canine TGF-beta 1 ELISA - Quantikine Best Seller
R&D Systems, part of Bio-Techne | Catalog # DB100C
Key Product Details
Assay Length
4.5 hours
Sample Type & Volume Required Per Well
Cell Culture Supernates (50 µL), Serum (10 µL), Platelet-poor Plasma (50 µL), Platelet-poor EDTA Plasma (50 µL), Urine (50 µL)
Sensitivity
5.5 pg/mL
Assay Range
31.3-2000 pg/mL (Cell Culture Supernates, Serum, Platelet-poor Plasma, Platelet-poor EDTA Plasma, Urine)
Product Summary for Human/Mouse/Rat/Porcine/Canine TGF-beta 1 Quantikine ELISA
The Quantikine™ Human/Mouse/Rat/Porcine/Canine TGF-beta 1 Immunoassay is a 4.5 hour solid phase ELISA designed to measure TGF-beta 1 in cell culture supernates, serum, platelet-poor plasma, and urine. It contains CHO cell-expressed recombinant TGF-beta 1 and has been shown to quantitate the recombinant factor accurately. Results obtained using natural 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 can be used to determine relative mass values for natural TGF-beta 1.
Product Specifications
Measurement
Quantitative ELISA
Detection Method
Colorimetric - 450nm (TMB)
Conjugate
HRP
Reactivity
Human, Mouse, Rat, Porcine, Canine
Specificity
Natural and recombinant TGF-beta 1. This assay also recognizes human TGF-beta 1.2.
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.
Sample Values
Serum/Plasma/Urine - Samples were evaluated for the presence of TGF-beta 1 in this assay.
*Only 40% of the urine samples meausered detectable levels (>31.3 pg/mL).
**Only 60% of the EDTA plasma samples measured detectable levels with the required 40 fold dilution. Two of five samples read just
below the standard curve (< 31.3 pg/mL).
| Human Samples | Mean (pg/mL) | Range (pg/mL) | Standard Devitation (pg/mL) |
| Serum (n=31) | 51,640 | 32,091-95,147 | 11,901 |
| platelet-poor EDTA plasma (n=31) | 2377 | 1414-4641 | 729 |
| Platelet-poor heparin plasma (n=31) | 2383 | 1445-3653 | 561 |
| Urine* (n=10) | 65.9 | ND-108 | --- |
ND=Non-detectable
| Mouse Samples | Mean (pg/mL) | Range (pg/mL) | Standard Deviation (pg/mL) |
| Serum (n=10) | 97,352 | 78,751-114,130 | 12,623 |
| Platelet-poor EDTA plasma (n=10) | 41,763 | 16,577-68,505 | 16,887 |
| Platelet-poor heparin plasma (n=10) | 41,101 | 18,195-66,700 | 17,419 |
| Rat Samples | Mean (pg/mL) | Range (pg/mL) | Standard Deviation (pg/mL) |
| Serum (n=10) | 68,108 | 43,721-89,138 | 14,343 |
| Platelet-poor EDTA plasma (n=5) | 5833 | 4912-7820 | 1141 |
| Platelet-poor heparin plasma (n=5) | 13,971 | 5274-26,114 | 8392 |
| Porcine Samples | Mean (pg/mL) | Range (pg/mL) | Standard Deviation (pg/mL) |
| Serum (n=5) | 17,597 | 13,481-23,914 | 4146 |
| Platelet-poor EDTA plasma (n=5) | 1955 | 1530-2810 | 518 |
| Platelet-poor heparin plasma (n=5) | 1567 | 1111-2475 | 551 |
| Canine Samples | Mean (pg/mL) | Range (pg/mL) | Standard Deviation (pg/mL) |
| Serum (n=10) | 33,455 | 2193-79,590 | 19,795 |
| Platelet-poor EDTA plasma** (n=5) | 2195 | ND-2515 | — |
| Platelet-poor heparin plasma*** (n=5) | 3763 | ND-4278 | — |
***Only 80% of the heparin plasma samples measured detectable levels with the required 40 fold dilution. One of five samples read
just below the standard curve (< 31.3 pg/mL).
SAMPLE VALUES CONTINUED
Cell Culture Supernates:
Cell Culture Supernates:
Human peripheral blood mononuclear cells (PBMCs) were separated from whole blood by a
density gradient centrifugation method using Ficoll-Paque Plus. CD4+ T cells were isolated from
PBMCs using the MagCellect™ Human CD4+ T cell Isolation Kit (R&D Systems®,
Catalog # MAGH102). Cells were seeded at 5 x 105
/mL and cultured using Excellerate™
Human T Cell Expansion Media, Xeno-Free (R&D Systems, Catalog # CCM030). T cells were left
untreated or treated with 10 ng/mL GMP recombinant human (rh) IL-7 (R&D Systems,
Catalog # 207-GMP), 10 ng/mL GMP rhIL-15 (R&D Systems, Catalog # 247-GMP), and stimulated
via their T cell receptor (TCR) and co-stimulatory receptor for 5 days. TCR stimulation was
mediated using 25 μL Cloudz™ CD3/28 particles (Cloudz T Cell Activation Kit - CD3/CD28,
(R&D Systems, Catalog # CLD001) per mL of culture media. CD4+ T cells were maintained in a
5% CO2
incubator at 37 °C for 5 days. An aliquot of the cell culture supernates was removed,
assayed for TGF-beta 1, and measured 77.7 pg/mL (untreated) and 471 pg/mL (treated).
Human PBMCs were seeded at 1 x 106
/mL and cultured in RPMI supplemented with 10% fetal
bovine serum (FBS), 2 mM L-glutamine, 100 U/mL penicillin, and 100 μg/mL streptomycin and
left untreated. An aliquot of the cell culture supernate was removed, assayed for TGF-beta 1, and
measured 2854 pg/mL.
Mouse EL-4 cells were cultured in DMEM High Glucose supplemented with 10% FBS,
2 mM L-glutamine, 100 U/mL penicillin, and 100 μg/mL streptomycin. Cells were then treated
with 10 ng/mL PMA and 10 ug/mL PHA for 24 hours. An aliquot of the cell culture supernate
was removed, assayed for TGF-beta 1, and measured 2969 pg/mL.
Rat spleen was taken from a pregnant Sprague Dawley rat, homogenized, and cultured in DME
with 10% FBS, 2 mM L-glutamine, 100 U/mL penicillin, and 100 μg/mL streptomycin at 37 °C
and 5% CO2
. Rat splenocytes were treated with 50 ng/mL recombinant rat IL-2 and 5 μg/mL
PHA for 3 days. An aliquot of the cell culture supernate was removed, assayed for TGF-beta 1, and
measured 2716 pg/mL.
Porcine PK-15 cells were cultured in MEM supplemented with 10% FBS, 2 mM L-glutamine,
100 U/mL penicillin, and 100 μg/mL streptomycin. Cells were treated with 1 μg/mL LPS for 72
hours. An aliquot of the cell culture supernate was removed, assayed for TGF-beta 1, and measured
1631 pg/mL.
Precision
Intra-Assay Precision (Precision within an assay) Three samples of known concentration were tested twenty times 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, Platelet-poor EDTA Plasma, Platelet-poor Plasma, Serum, Urine
| Intra-Assay Precision | Inter-Assay Precision | |||||||||||
|---|---|---|---|---|---|---|---|---|---|---|---|---|
| Sample | 1 | 2 | 3 | 4 | 5 | 6 | 1 | 2 | 3 | 4 | 5 | 6 |
| n | 20 | 20 | 20 | 20 | 20 | 20 | 20 | 20 | 20 | 20 | 20 | 20 |
| Mean (pg/mL) | 317 | 683 | 1271 | 309 | 708 | 1072 | 312 | 657 | 1184 | 303 | 622 | 1092 |
| Standard Deviation | 11.3 | 42.2 | 31.1 | 15.3 | 40.0 | 44.3 | 26.1 | 46.6 | 92.3 | 21.2 | 47.2 | 70.0 |
| CV% | 3.6 | 6.2 | 2.4 | 5.0 | 5.6 | 4.1 | 8.4 | 7.1 | 7.8 | 7.0 | 7.6 | 6.4 |
Recovery for Human/Mouse/Rat/Porcine/Canine TGF-beta 1 Quantikine ELISA
The recovery of TGF-beta 1 spiked to levels throughout the range of the assay in activated samples was evaluated.
| Sample Type | Average % Recovery | Range % |
|---|---|---|
| Media + FBS (n=4) | 103 | 80-125 |
| Platelet-poor EDTA Plasma (n=4) | 90 | 83-98 |
| Platelet-poor Plasma (n=4) | 90 | 77-107 |
| Serum-free Cell Culture Media (n=4) | 87 | 82-95 |
| Urine (n=4) | 114 | 105-123 |
Linearity
To assess linearity of the assay, the activated samples containing and/or spiked with high concentrations of TGF-beta 1, were diluted with calibrator diluent to produce samples with values within the dynamic range of the assay.
Scientific Data Images for Human/Mouse/Rat/Porcine/Canine TGF-beta 1 Quantikine ELISA
Human TGF-beta 1 ELISA Cell culture supernate/Urine Standard Curve
Human TGF-beta 1 ELISA Serum/Platelet-poor Plasma 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: TGF-beta 1
Transforming Growth Factor Beta 1, 2, and 3 (TGF-beta 1, TGF-beta 2, and TGF-beta 3) are highly pleiotropic cytokines that virtually all cell types secrete. TGF-beta molecules are proposed to act as cellular switches that regulate processes such as immune function, proliferation, and epithelial-mesenchymal transition. Targeted deletions of these genes in mice show that each TGF-beta isoform has some non-redundant functions: TGF-beta 1 is involved in hematopoiesis and endothelial differentiation; TGF-beta 2 affects development of cardiac, lung, craniofacial, limb, eye, ear, and urogenital systems; and TGF-beta 3 influences palatogenesis and pulmonary development. The full range of in vitro biological activities of TGF-beta 5 has not yet been explored. However, TGF-beta 1, TGF-beta 2, TGF-beta 3, and TGF-beta 5 have been found to be largely interchangeable in an inhibitory bioassay, and it is anticipated that TGF-beta 5 will show a spectrum of activities similar to the other TGF-beta family members. To date, the production of TGF-beta 5 has only been demonstrated in Xenopus.
TGF-beta ligands are initially synthesized as precursor proteins that undergo proteolytic cleavage. The mature segments form active ligand dimers via a disulfide-rich core consisting of the characteristic 'cysteine knot'. TGF-beta signaling begins with binding to a complex of the accessory receptor betaglycan (also known as TGF-beta RIII) and a type II serine/threonine kinase receptor termed TGF-beta RII. This receptor then phosphorylates and activates a type I serine/threonine kinase receptor, either ALK-1 or TGF-beta RI (also called ALK-5). The activated type I receptor phosphorylates and activates Smad proteins that regulate transcription. Use of other signaling pathways that are Smad-independent allows for distinct actions observed in response to TGF-beta in different contexts.
Long Name
Transforming Growth Factor beta 1
Alternate Names
TGF beta1, TGFB, TGFB1, TGFbeta 1
Gene Symbol
TGFB1
Additional TGF-beta 1 Products
Product Documents for Human/Mouse/Rat/Porcine/Canine TGF-beta 1 Quantikine ELISA
Product Specific Notices for Human/Mouse/Rat/Porcine/Canine TGF-beta 1 Quantikine ELISA
For research use only
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