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Human/Mouse/Rat/Porcine/Canine TGF-beta 1 ELISA - Quantikine Best Seller

Bio-Techne includes R&D Systems | Catalog # DB100C

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DB100C
PDB100C
SB100C

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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

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.

Human SamplesMean (pg/mL)Range (pg/mL)Standard Devitation (pg/mL)
Serum (n=31)51,64032,091-95,14711,901
platelet-poor EDTA plasma (n=31)23771414-4641729
Platelet-poor heparin plasma (n=31)23831445-3653561
Urine* (n=10)65.9ND-108---
*Only 40% of the urine samples meausered detectable levels (>31.3 pg/mL).
ND=Non-detectable

Mouse SamplesMean (pg/mL)Range (pg/mL)Standard Deviation (pg/mL)
Serum (n=10)97,35278,751-114,13012,623
Platelet-poor EDTA plasma (n=10)41,76316,577-68,50516,887
Platelet-poor heparin plasma (n=10)41,10118,195-66,70017,419

Rat SamplesMean (pg/mL)Range (pg/mL)Standard Deviation (pg/mL)
Serum (n=10)68,10843,721-89,13814,343
Platelet-poor EDTA plasma (n=5)58334912-78201141
Platelet-poor heparin plasma (n=5)13,9715274-26,1148392

Porcine SamplesMean (pg/mL)Range (pg/mL)Standard Deviation (pg/mL)
Serum (n=5)17,59713,481-23,9144146
Platelet-poor EDTA plasma (n=5)19551530-2810518
Platelet-poor heparin plasma (n=5)15671111-2475551

Canine SamplesMean (pg/mL)Range (pg/mL)Standard Deviation (pg/mL)
Serum (n=10)33,4552193-79,59019,795
Platelet-poor EDTA plasma** (n=5)2195ND-2515
Platelet-poor heparin plasma*** (n=5)3763ND-4278
**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).

***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:
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.

Human TGF-beta 1 ELISA Linearity ELISA

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 Cell culture supernate/Urine Standard Curve

Human TGF-beta 1 ELISA Serum/Platelet-poor Plasma 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

Entrez Gene IDs

7040 (Human); 21803 (Mouse); 59086 (Rat); 397078 (Porcine); 100033900 (Equine)

Gene Symbol

TGFB1

Additional TGF-beta 1 Products

Product Documents for Human/Mouse/Rat/Porcine/Canine TGF-beta 1 Quantikine ELISA

Certificate of Analysis

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Note: Certificate of Analysis not available for kit components.

Product Specific Notices for Human/Mouse/Rat/Porcine/Canine TGF-beta 1 Quantikine ELISA

For research use only

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Customer Reviews for Human/Mouse/Rat/Porcine/Canine TGF-beta 1 Quantikine ELISA (2)

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  • Human/Mouse/Rat/Porcine/Canine TGF-beta 1 Quantikine ELISA
    Name: Michiaki Mukai
    Sample Tested: Platelet-rich plasma
    Verified Customer | Posted 02/25/2024
  • Human/Mouse/Rat/Porcine/Canine TGF-beta 1 Quantikine ELISA
    Name: Fumou Sun
    Sample Tested: Serum and Plasma
    Verified Customer | Posted 05/26/2023
    The TGF-beta 1 Quantikine ELISA Kit is stable. The R2 value of the standard curve is more than 0.99. Easy to use!
    Human/Mouse/Rat/Porcine/Canine TGF-beta 1 Quantikine ELISA DB100C

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