Human TGF-beta 2 DuoSet ELISA

Catalog # Availability Size / Price Qty
DY302
Ancillary Products Available
Human TGF-beta 2 ELISA Standard Curve
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Product Details
Procedure
Citations (12)
FAQs
Supplemental Products
Reviews

Human TGF-beta 2 DuoSet ELISA Summary

Assay Type
Solid Phase Sandwich ELISA
Format
96-well strip plate
Sample Volume Required
100 µL
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)

 

  

Data Example

Human TGF-beta 2 ELISA Standard Curve

Product Datasheets

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

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 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.

12 Citations: Showing 1 - 10
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  1. 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
  2. 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
  3. 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
  4. 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
  5. 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
  6. 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
  7. 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
  8. 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
  9. 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
  10. 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
  11. 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
  12. 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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