Human TGF-beta 2 Quantikine ELISA Kit

R&D Systems | Catalog # DB250

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Key Product Details

Assay Length

4.5 hours

Sample Type & Volume Required Per Well

Cell Culture Supernates (125 µL), Serum (125 µL), EDTA Plasma (125 µL), Heparin Plasma (125 µL), Citrate Plasma (125 µL)

Sensitivity

7 pg/mL

Assay Range

31.2-2000 pg/mL (Cell Culture Supernates, Serum, EDTA Plasma, Heparin Plasma, Citrate Plasma)
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Product Summary for Human TGF-beta 2 Quantikine ELISA Kit

The Quantikine Human TGF-beta 2 Immunoassay is a 4.5 hour solid-phase ELISA designed to measure activated TGF-beta 2 in cell culture supernates, serum, and plasma. The Quantikine Human TGF-beta 2 Immunoassay contains CHO cell-derived recombinant human TGF-beta 2 and antibodies raised against the recombinant protein. The assay has been shown to quantitate the recombinant factor accurately. Results obtained using human TGF-beta 2 showed linear curves that were parallel to the standard curves obtained using the recombinant TGF-beta 2 kit standards. These results indicate that this kit can be used to determine relative mass values for natural human TGF-beta 2.

Product Specifications

Assay Type

Solid Phase Sandwich ELISA

Format

96-well strip plate

Measurement

Quantitative ELISA

Detection Method

Colorimetric - 450nm (TMB)

Conjugate

HRP

Species

Human

Specificity

Natural and recombinant human TGF-beta 2 and natural porcine TGF-beta 2.

Cross-reactivity

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

Interference

Interference observed with 1 or more 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.

Cell Culture Supernates, Citrate Plasma, EDTA Plasma, Heparin Plasma, Serum

Intra-Assay Precision Inter-Assay Precision
Sample 1 2 3 1 2 3
n 20 20 20 20 20 20
Mean (pg/mL) 194 509 1044 196 500 1015
Standard Deviation 13.8 27.0 28.3 9.8 21.5 43.6
CV% 7.1 5.3 2.7 5.0 4.3 4.3

Recovery for Human TGF-beta 2 Quantikine ELISA Kit

To assess recovery of the assay, samples of various matrices were spiked with three concentrations of TGF-beta 2, activated, and measured in the assay.

Sample Type Average % Recovery Range %
Cell Culture Media (n=5) 101 89-113
Citrate Plasma (n=5) 107 93-116
EDTA Plasma (n=5) 101 95-111
Heparin Plasma (n=5) 102 89-113
Serum (n=5) 86 73-95

Linearity

To assess the linearity of the assay, samples were spiked with high concentrations of TGF-beta 2, activated and then serially diluted with Calibrator Diluent to produce samples with values within the dynamic range of the assay.

Human TGF-beta 2 ELISA Linearity

Scientific Data Images for Human TGF-beta 2 Quantikine ELISA Kit

Human TGF-beta 2 ELISA Standard Curve

Human TGF-beta 2 ELISA Standard Curve

Detection of Human TGF-beta 2 by ELISA

Detection of Human TGF-beta 2 by ELISA

ELISA for active and total TGF-beta 2 in the media (A, B). At day 22, the concentration of active and total TGF-beta 2 in the media of IL-13 pre-treated NHBE at 1 and 10 ng/ml is significantly higher as compared to untreated NHBE (0 ng/ml of IL-13) media; * p < 0.01. At day 22 and day 28, the concentration of active TGF-beta 2 in the IL-13 pre-treated NHBE at 10 ng/ml is elevated compared to pre-treated NHBE at 1 ng/ml; # p < 0.01. At day 28 active and total TGF-beta 2 in IL-13 pre-treated NHBE at 10 ng/ml is increased compared to untreated NHBE; * p < 0.01. (C, D) At day 25, the NHBE pre-treated with IL-13 at 10 ng/ml, has higher levels of active and total TGF-beta 2 in the media as compared to untreated and pre-treated NHBE at 1 ng/ml co-cultured with NHLF (*, # p < 0.01 compared to 0 and 1 ng/ml IL-13 pre-treated NHBE co-cultured with NHLF, respectively). At day 31, there is no significant difference in the levels of active and total TGF-beta 2 between treatment conditions. NHLF represents levels of active and total TGF-beta 2 in media of fibroblasts in a collagen gel without NHBE co-culture. All experiments were performed using 3 donors, grown in duplicate, with 3–6 wells per condition. Image collected and cropped by CiteAb from the following open publication (https://respiratory-research.biomedcentral.com/articles/10.1186/1465-99…), licensed under a CC-BY license. Not internally tested by R&D Systems.
Detection of Human TGF-beta 2 by ELISA

Detection of Human TGF-beta 2 by ELISA

ELISA for active and total TGF-beta 2 in the media (A, B). At day 22, the concentration of active and total TGF-beta 2 in the media of IL-13 pre-treated NHBE at 1 and 10 ng/ml is significantly higher as compared to untreated NHBE (0 ng/ml of IL-13) media; * p < 0.01. At day 22 and day 28, the concentration of active TGF-beta 2 in the IL-13 pre-treated NHBE at 10 ng/ml is elevated compared to pre-treated NHBE at 1 ng/ml; # p < 0.01. At day 28 active and total TGF-beta 2 in IL-13 pre-treated NHBE at 10 ng/ml is increased compared to untreated NHBE; * p < 0.01. (C, D) At day 25, the NHBE pre-treated with IL-13 at 10 ng/ml, has higher levels of active and total TGF-beta 2 in the media as compared to untreated and pre-treated NHBE at 1 ng/ml co-cultured with NHLF (*, # p < 0.01 compared to 0 and 1 ng/ml IL-13 pre-treated NHBE co-cultured with NHLF, respectively). At day 31, there is no significant difference in the levels of active and total TGF-beta 2 between treatment conditions. NHLF represents levels of active and total TGF-beta 2 in media of fibroblasts in a collagen gel without NHBE co-culture. All experiments were performed using 3 donors, grown in duplicate, with 3–6 wells per condition. Image collected and cropped by CiteAb from the following open publication (https://respiratory-research.biomedcentral.com/articles/10.1186/1465-99…), licensed under a CC-BY license. Not internally tested by R&D Systems.
Detection of Human TGF-beta 2 by ELISA

Detection of Human TGF-beta 2 by ELISA

ELISA for active and total TGF-beta 2 in the media (A, B). At day 22, the concentration of active and total TGF-beta 2 in the media of IL-13 pre-treated NHBE at 1 and 10 ng/ml is significantly higher as compared to untreated NHBE (0 ng/ml of IL-13) media; * p < 0.01. At day 22 and day 28, the concentration of active TGF-beta 2 in the IL-13 pre-treated NHBE at 10 ng/ml is elevated compared to pre-treated NHBE at 1 ng/ml; # p < 0.01. At day 28 active and total TGF-beta 2 in IL-13 pre-treated NHBE at 10 ng/ml is increased compared to untreated NHBE; * p < 0.01. (C, D) At day 25, the NHBE pre-treated with IL-13 at 10 ng/ml, has higher levels of active and total TGF-beta 2 in the media as compared to untreated and pre-treated NHBE at 1 ng/ml co-cultured with NHLF (*, # p < 0.01 compared to 0 and 1 ng/ml IL-13 pre-treated NHBE co-cultured with NHLF, respectively). At day 31, there is no significant difference in the levels of active and total TGF-beta 2 between treatment conditions. NHLF represents levels of active and total TGF-beta 2 in media of fibroblasts in a collagen gel without NHBE co-culture. All experiments were performed using 3 donors, grown in duplicate, with 3–6 wells per condition. Image collected and cropped by CiteAb from the following open publication (https://respiratory-research.biomedcentral.com/articles/10.1186/1465-99…), licensed under a CC-BY license. Not internally tested by R&D Systems.
Detection of Human TGF-beta 2 by ELISA

Detection of Human TGF-beta 2 by ELISA

ELISA for active and total TGF-beta 2 in the media (A, B). At day 22, the concentration of active and total TGF-beta 2 in the media of IL-13 pre-treated NHBE at 1 and 10 ng/ml is significantly higher as compared to untreated NHBE (0 ng/ml of IL-13) media; * p < 0.01. At day 22 and day 28, the concentration of active TGF-beta 2 in the IL-13 pre-treated NHBE at 10 ng/ml is elevated compared to pre-treated NHBE at 1 ng/ml; # p < 0.01. At day 28 active and total TGF-beta 2 in IL-13 pre-treated NHBE at 10 ng/ml is increased compared to untreated NHBE; * p < 0.01. (C, D) At day 25, the NHBE pre-treated with IL-13 at 10 ng/ml, has higher levels of active and total TGF-beta 2 in the media as compared to untreated and pre-treated NHBE at 1 ng/ml co-cultured with NHLF (*, # p < 0.01 compared to 0 and 1 ng/ml IL-13 pre-treated NHBE co-cultured with NHLF, respectively). At day 31, there is no significant difference in the levels of active and total TGF-beta 2 between treatment conditions. NHLF represents levels of active and total TGF-beta 2 in media of fibroblasts in a collagen gel without NHBE co-culture. All experiments were performed using 3 donors, grown in duplicate, with 3–6 wells per condition. Image collected and cropped by CiteAb from the following open publication (https://respiratory-research.biomedcentral.com/articles/10.1186/1465-99…), licensed under a CC-BY license. Not internally tested by R&D Systems.

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 2

TGF-beta 2 is synthesized as a prepro-cytokine with a 19 amino acid (aa) signal sequence, a 283 aa pro-region, and a 112 aa mature segment (1-5). It dimerizes with formation of disulfide bonds between the 'pro' regions and disulfide bonds between the 'mature' regions. The mature region is 71% and 80% identical with human TGF-beta 1 and TGF-beta 3 (6, 7) and 97% identical with the corresponding mouse protein (8). After proteolytic cleavage of the disulfide-linked mature region, it remains hydrogen-bonded to the disulfide-linked prosegments (LAP or latencyassociated protein) (1, 2, 9). If secreted in this form, LAP keeps TGF-beta 2 in an inactive state until dissociation, caused by proteases, glycosidases, or extreme pH (2, 9). In many types of cells, an additional protein, latent TGF-beta binding protein (LTBP), is covalently linked to the LAP homodimer prior to secretion. LTBP, a 130 kDa cysteine-rich glycoprotein, creates a 235 kDa large latent complex that is secreted, most likely binding to the extracellular matrix (1, 9-11). The latency components are believed to act as natural antagonists of TGF-beta activity, to target TGF-beta to distinct tissues, and to maintain a reservoir of TGF-beta (1, 2, 12). On release from latency, active homodimeric TGF-beta can bind to cell-surface receptors or to other proteins, such as alpha 2-macroglobulin (13). 
The signal transducing receptor for TGF-beta 2 is a heterotetrameric complex of two type I signal-transducing receptors (53 kDa; TGF-beta RI) and two type II ligand-binding receptors (75-85 kDa; TGF-beta RII) (14-19). The binding of TGF-beta 2 appears to initially involve a type III TGF-beta receptor, either 300 kDa betaglycan (20) or 180 kDa endoglin (14, 21), which then "hands off" to TGF-beta RII. 
TGF-beta 2 is expressed by a variety of cells, including osteoclasts, thymic epithelium, keratinocytes, hepatocytes, chief cells of the stomach, satellite cells, skeletal muscle cells, prostatic epithelium, bronchial epithelium, neurons and astrocytes, fibroblasts and visceral smooth muscle, and macrophages (14-19). TGF-beta 2 has marked cross-species bioactivity (e.g., human TGF-beta 2 is active on mouse cells (33), while porcine TGF-beta 2 is active on rabbit cells (34)). TGF-beta 2 has four fundamental activities: it is a growth inhibitor for most types of cells; it enhances the deposition of extracellular matrix; it is immunosuppressive, suppressing APC expression of both IL-12 and CD40L while upregulating IL-10 secretion; and during fetal development, it is expressed in discrete areas, such as epithelium, myocardium, cartilage and bone of extremities and in the nervous system, suggesting specific functions (1, 35-37).

Long Name

Transforming Growth Factor beta 2

Alternate Names

TGFB2, TGFbeta 2

Entrez Gene IDs

7042 (Human); 21808 (Mouse); 397084 (Porcine)

Gene Symbol

TGFB2

Additional TGF-beta 2 Products

Product Documents for Human TGF-beta 2 Quantikine ELISA Kit

Certificate of Analysis

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Note: Certificate of Analysis not available for kit components.

Product Specific Notices for Human TGF-beta 2 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 TGF-beta 2 Quantikine ELISA Kit

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  • Human TGF-beta 2 Quantikine ELISA Kit
    Name: Alicia Furlan
    Sample Tested: EDTA Plasma and Serum
    Verified Customer | Posted 05/21/2024
  • Human TGF-beta 2 Quantikine ELISA Kit
    Name: Anonymous
    Sample Tested: Feline serum and Feline aqueous humor
    Verified Customer | Posted 04/01/2024
    Human TGF-beta 2 Quantikine ELISA Kit was used to quantify TGF-beta 2 in aqueous humor samples from cats with glaucoma.
    Human TGF-beta 2 Quantikine ELISA Kit DB250
  • Human TGF-beta 2 Quantikine ELISA Kit
    Name: Adam Guess
    Sample Tested: Cell Culture Supernates
    Verified Customer | Posted 02/04/2017
    Human TGF-beta 2 Quantikine ELISA Kit DB250

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Protocols

View specific protocols for Human TGF-beta 2 Quantikine ELISA Kit (DB250):

Refer to the product for complete assay procedure.

Bring all reagents and samples to room temperature before use. It is recommended that all samples, standards, and controls be assayed in duplicate.
  1.   Prepare all reagents, standard dilutions, and samples as directed in the product insert.
  2.   Remove excess microplate strips from the plate frame, return them to the foil pouch containing the desiccant pack, and reseal.
    3. 100 µL Assay Diluent
  3.   Add 100 µL of the appropriate Assay Diluent to each well.
    4. 100 µL Standard, Control, or Activated Sample
  4.   Add 100 µL of Standard, control, or activated sample to each well. Cover with a plate sealer, and incubate at room temperature for 2 hours.
  5.   Aspirate each well and wash, repeating the process twice for a total of 3 washes.
    6. 200 µL Conjugate
  6.   Add 200 µL of Conjugate to each well. Cover with a new plate sealer, and incubate at room temperature for 2 hours.
  7.   Aspirate and wash 3 times.
    8. 200 µL Substrate Solution
  8.   Add 200 µL Substrate Solution to each well. Incubate at room temperature for 20 minutes. PROTECT FROM LIGHT.
    9. 50 µL Stop Solution
  9.   Add 50 µL of Stop Solution to each well. Read at 450 nm within 30 minutes. Set wavelength correction to 540 nm or 570 nm.

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