Human TGF-beta 2 DuoSet ELISA

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Citations (18)
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Human TGF-beta 2 DuoSet ELISA Summary

Assay Type
Solid Phase Sandwich ELISA
Format
96-well strip plate
Sample Volume Required
100 µL
Assay Range
31.2 - 2,000 pg/mL
Sufficient Materials
For fifteen 96-well plates*
Specificity
Please see the product datasheet

* Provided that the recommended microplates, buffers, diluents, substrates and solutions are used, and the assay is run as summarized in the Assay Procedure provided.

This DuoSet ELISA Development kit contains the basic components required for the development of sandwich ELISAs to measure natural and recombinant human TGF-beta2. The suggested diluent is suitable for the analysis of most cell culture supernate samples. Diluents for complex matrices, such as serum and plasma, should be evaluated prior to use in this DuoSet.

 

Product Features

  • Optimized capture and detection antibody pairings with recommended concentrations save lengthy development time
  • Development protocols are provided to guide further assay optimization
  • Assay can be customized to your specific needs
  • Economical alternative to complete kits

Kit Content

  • Capture Antibody
  • Detection Antibody
  • Recombinant Standard
  • Streptavidin conjugated to horseradish-peroxidase (Streptavidin-HRP)

Other Reagents Required

DuoSet Ancillary Reagent Kit 2 (5 plates): (Catalog # DY008) containing 96 well microplates, plate sealers, substrate solution, stop solution, plate coating buffer (PBS), wash buffer, and Reagent Diluent Concentrate 2.

The components listed above may be purchased separately:

PBS: (Catalog # DY006), or 137 mM NaCl, 2.7 mM KCl, 8.1 mM Na2HPO4, 1.5 mM KH2PO4, pH 7.2 - 7.4, 0.2 µm filtered

Wash Buffer: (Catalog # WA126), or 0.05% Tween® 20 in PBS, pH 7.2-7.4

Reagent Diluent: (Catalog # DY995), or 1% BSA in PBS, pH 7.2-7.4, 0.2 µm filtered

Substrate Solution: 1:1 mixture of Color Reagent A (H2O2) and Color Reagent B (Tetramethylbenzidine) (Catalog # DY999)

Stop Solution: 2 N H2SO4 (Catalog # DY994)

Microplates: R&D Systems (Catalog # DY990)

Plate Sealers: ELISA Plate Sealers (Catalog # DY992)

Scientific Data

Human TGF-beta 2 ELISA Standard Curve

Product Datasheets

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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
Entrez Gene IDs:
7042 (Human); 21808 (Mouse); 397084 (Porcine)
Alternate Names:
BSC-1 cell growth inhibitor; cetermin; Glioblastoma-derived T-cell suppressor factor; G-TSF; MGC116892; polyergin; TGFB2; TGFbeta 2; TGF-beta 2; TGF-beta2; TGF-beta-2; transforming growth factor beta-2; transforming growth factor, beta 2

Assay Procedure

GENERAL ELISA PROTOCOL

Plate Preparation

  1. Dilute the Capture Antibody to the working concentration in PBS without carrier protein. Immediately coat a 96-well microplate with 100 μL per well of the diluted Capture Antibody. Seal the plate and incubate overnight at room temperature.
  2. Aspirate each well and wash with Wash Buffer, repeating the process two times for a total of three washes. Wash by filling each well with Wash Buffer (400 μL) using a squirt bottle, manifold dispenser, or autowasher. Complete removal of liquid at each step is essential for good performance. After the last wash, remove any remaining Wash Buffer by aspirating or by inverting the plate and blotting it against clean paper towels.
  3. Block plates by adding 300 μL Reagent Diluent to each well. Incubate at room temperature for a minimum of 1 hour.
  4. Repeat the aspiration/wash as in step 2. The plates are now ready for sample addition.

Assay Procedure

  1. Add 100 μL of sample or standards in Reagent Diluent, or an appropriate diluent, per well. Cover with an adhesive strip and incubate 2 hours at room temperature.
  2. Repeat the aspiration/wash as in step 2 of Plate Preparation.
  3. Add 100 μL of the Detection Antibody, diluted in Reagent Diluent, to each well. Cover with a new adhesive strip and incubate 2 hours at room temperature.
  4. Repeat the aspiration/wash as in step 2 of Plate Preparation.
  5. Add 100 μL of the working dilution of Streptavidin-HRP to each well. Cover the plate and incubate for 20 minutes at room temperature. Avoid placing the plate in direct light.
  6. Repeat the aspiration/wash as in step 2.
  7. Add 100 μL of Substrate Solution to each well. Incubate for 20 minutes at room temperature. Avoid placing the plate in direct light.
  8. Add 50 μL of Stop Solution to each well. Gently tap the plate to ensure thorough mixing.
  9. Determine the optical density of each well immediately, using a microplate reader set to 450 nm. If wavelength correction is available, set to 540 nm or 570 nm. If wavelength correction is not available, subtract readings at 540 nm or 570 nm from the readings at 450 nm. This subtraction will correct for optical imperfections in the plate. Readings made directly at 450 nm without correction may be higher and less accurate.

 

Citations for Human TGF-beta 2 DuoSet ELISA

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.

18 Citations: Showing 1 - 10
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  1. Role of Betaglycan in TGF-beta Signaling and Wound Healing in Human Endometriotic Epithelial Cells and in Endometriosis
    Authors: AN Mwaura, MA Riaz, JB Maoga, E Mecha, COA Omwandho, G Scheiner-B, I Meinhold-H, L Konrad
    Biology, 2022;11(4):.
    Species: Human
    Sample Types: Cell Culture Supernates
  2. Nucleoporin-93 reveals a common feature of aggressive breast cancers: robust nucleocytoplasmic transport of transcription factors
    Authors: NB Nataraj, A Noronha, JS Lee, S Ghosh, HR Mohan Raju, A Sekar, B Zuckerman, M Lindzen, E Tarcitano, S Srivastava, M Selitrenni, I Livneh, D Drago-Garc, O Rueda, C Caldas, S Lev, T Geiger, A Ciechanove, I Ulitsky, R Seger, E Ruppin, Y Yarden
    Cell Reports, 2022;38(8):110418.
    Species: Human
    Sample Types: Cell Culture Supernates
  3. Csnk1a1 inhibition modulates the inflammatory secretome and enhances response to radiotherapy in glioma
    Authors: G Liu, H Li, W Zhang, J Yu, X Zhang, R Wu, M Niu, X Liu, R Yu
    Journal of Cellular and Molecular Medicine, 2021;0(0):.
    Species: Human
    Sample Types: Cell Culture Supernates
  4. Latent TGFbeta-binding proteins regulate UCP1 expression and function via TGFbeta2
    Authors: D Halbgebaue, J Roos, JB Funcke, H Neubauer, BS Hamilton, E Simon, EZ Amri, KM Debatin, M Wabitsch, P Fischer-Po, D Tews
    Molecular Metabolism, 2021;53(0):101336.
    Species: Human
    Sample Types: Cell Culture Supernates
  5. Regulation of Fibroblast Activation Protein by Transforming Growth Factor Beta-1 in Glioblastoma Microenvironment
    Authors: E Krepela, Z Vanickova, P Hrabal, M Zubal, B Chmielova, E Balaziova, P Vymola, I Matrasova, P Busek, A Sedo
    International Journal of Molecular Sciences, 2021;22(3):.
    Species: Human
    Sample Types: Cell Lysates
  6. Mesenchymal Stem and Stromal Cells Harness Macrophage-Derived Amphiregulin to Maintain Tissue Homeostasis
    Authors: JH Ko, HJ Kim, HJ Jeong, HJ Lee, JY Oh
    Cell Rep, 2020;30(11):3806-3820.e6.
    Species: Human
    Sample Types: Cell Culture Supernates
  7. Inhibition of IRE1 RNase activity modulates the tumor cell secretome and enhances response to chemotherapy
    Authors: SE Logue, EP McGrath, P Cleary, S Greene, K Mnich, A Almanza, E Chevet, RM Dwyer, A Oommen, P Legembre, F Godey, EC Madden, B Leuzzi, J Obacz, Q Zeng, JB Patterson, R Jäger, AM Gorman, A Samali
    Nat Commun, 2018;9(1):3267.
    Species: Human
    Sample Types: Cell Culture Supernates
  8. Altered Decorin Biology in Proliferative Vitreoretinopathy: A Mechanistic and Cohort Study
    Authors: G Begum, J O'Neill, R Chaudhary, K Blachford, DRJ Snead, M Berry, RAH Scott, A Logan, RJ Blanch
    Invest. Ophthalmol. Vis. Sci., 2018;59(12):4929-4936.
    Species: Human
    Sample Types: Vitreous Humor
  9. Asthmatic airway epithelial cells differentially regulate fibroblast expression of extracellular matrix components.
    Authors: Reeves S, Kolstad T, Lien T, Elliott M, Ziegler S, Wight T, Debley J
    J Allergy Clin Immunol, 2014;134(3):663-670.e1.
    Species: Human
    Sample Types: Cell Culture Supernates
  10. Activin A as a mediator of NK-dendritic cell functional interactions.
    Authors: Seeger P, Bosisio D, Parolini S, Badolato R, Gismondi A, Santoni A, Sozzani S
    J Immunol, 2014;192(3):1241-8.
    Species: Human
    Sample Types: Cell Culture Supernates
  11. Interaction with colon cancer cells hyperactivates TGF-beta signaling in cancer-associated fibroblasts.
    Authors: Hawinkels L, Paauwe M, Verspaget H, Wiercinska E, van der Zon J, van der Ploeg K, Koelink P, Lindeman J, Mesker W, ten Dijke P, Sier C
    Oncogene, 2014;33(1):97-107.
    Species: Human
    Sample Types: Cell Culture Supernates
  12. Primary culture of avian embryonic heart forming region cells to study the regulation of vertebrate early heart morphogenesis by vitamin A.
    Authors: Cakstina I, Riekstina U, Boroduskis M, Nakurte I, Ancans J, Zile M, Muiznieks I
    BMC Dev Biol, 2014;14(0):10.
    Species: Chicken
    Sample Types: Cell Culture Supernates
  13. The peritoneum is both a source and target of TGF-beta in women with endometriosis.
    Authors: Young V, Brown J, Saunders P, Duncan W, Horne A
    PLoS ONE, 2014;9(9):e106773.
    Species: Human
    Sample Types: Peritoneal Fluid
  14. Glioblastoma-secreted factors induce IGFBP7 and angiogenesis by modulating Smad-2-dependent TGF-beta signaling.
    Authors: Pen A, Moreno MJ, Durocher Y, Deb-Rinker P, Stanimirovic DB
    Oncogene, 2008;27(54):6834-44.
    Species: Human
    Sample Types: Cell Culture Supernates
  15. Transforming growth factor-beta1 suppresses airway hyperresponsiveness in allergic airway disease.
    Authors: Alcorn JF, Rinaldi LM, Jaffe EF, van Loon M, Bates JH, Janssen-Heininger YM, Irvin CG
    Am. J. Respir. Crit. Care Med., 2007;176(10):974-82.
    Species: Mouse
    Sample Types: BALF
  16. Opposite regulation of transforming growth factors-beta2 and -beta3 expression in the human endometrium.
    Authors: Gaide Chevronnay HP, Cornet PB, Delvaux D, Lemoine P, Courtoy PJ, Henriet P, Marbaix E
    Endocrinology, 2007;149(3):1015-25.
    Species: Human
    Sample Types: Tissue Homogenates
  17. Activation of transforming growth factor-beta by the integrin alphavbeta8 delays epithelial wound closure.
    Authors: Neurohr C, Nishimura SL, Sheppard D
    Am. J. Respir. Cell Mol. Biol., 2006;35(2):252-9.
    Species: Human
    Sample Types: Cell Culture Supernates
  18. The selective estrogen receptor modulator arzoxifene and the rexinoid LG100268 cooperate to promote transforming growth factor beta-dependent apoptosis in breast cancer.
    Authors: Rendi MH, Suh N, Lamph WW, Krajewski S, Reed JC, Heyman RA, Berchuck A, Liby K, Risingsong R, Royce DB, Williams CR, Sporn MB
    Cancer Res., 2004;64(10):3566-71.
    Species: Rat
    Sample Types: Cell Culture Supernates

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