Human Latent TGF-beta 1 ELISpot Development Module, 5 Plate

Catalog # Availability Size / Price Qty
SEL246
Product Details
Citations (1)
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Human Latent TGF-beta 1 ELISpot Development Module, 5 Plate Summary

Assay Type
Quantitative Sandwich ELISA Development Module
Format
96-well microplate, sold separately (see Other Reagents Required)
Assay Length
3 hours 35 mins to 4 hours 50 mins**
Sample Type
Whole Cells
Sufficient Materials
For five 96-well microplates
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.


PRODUCT SUMMARY
Complete ELISpot kits are highly sensitive, microplate-based assays for the detection of cytokine secreting cells. Kits are available for detection and enumeration of a single analyte or two analytes simultaneously. Complete ELISpot kits are ready-to-run and require no assay development or refinement. ELISpot Development Modules contain the basic components required to develop an ELISpot assay. They offer an economical alternative to buying separate antibodies.

ELISpot development modules are an alternative to ELISpot kits. A basic understanding of ELISpot assay development is required for the successful use of these reagents. Each investigator should optimize the coating conditions, the assay sensitivity, the type of enzyme and substrate, as well as the concentrations of the capture and detection antibodies to achieve desired results. The analyte-specific ELISpot Development Module and the ELISpot Blue Color Module contain the necessary components for analyte detection and visualization, respectively. These modules can be used together but are sold separately. Each module contains enough reagents for at least five 96-well microplates.


PRODUCT FEATURES

  • An economical alternative to ELISpot Kits
  • Optimized capture and detection antibody pairings and recommended concentrations save lengthy development time
  • Generic development protocols provide direction to start an optimization protocol
  • Customize the assay to your specific needs


MODULE CONTENTS

  • Human TGF-beta 1 Capture Antibody
  • Human TGF-beta 1 Biotinylated-conjugated Detection Antibody


OTHER REAGENTS REQUIRED

  • ELISpot Blue Color Module or equivalent (R&D Systems, Catalog # SEL002)
  • PBS - 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 - 0.05% Tween® 20 in PBS
  • Blocking Buffer - 1% BSA, 5% Sucrose in PBS
  • Reagent Diluent - 1% BSA in PBS, pH 7.2 - 7.4, 0.2 µm filtered
  • 2 °C – 8 °C refrigerator
  • 37 °C CO2 incubator
  • Positive Control - Use Recombinant Human TGF-beta 1 or cells known to secrete Human TGF-beta 1
  • 96-well plates - Nitrocellulose-bottom plates, PVDF-bottom Immunospot® plates, or flat-bottom polystyrene Immulon® ELISA plates
  • Multi-channel pipette, squirt bottle, manifold dispenser, or automated microplate washer
  • Dissection microscope or an automated ELISpot Reader
  • Deionized H2O

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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 -20 to -70 °C. Use a manual defrost freezer and avoid repeated freeze-thaw cycles. 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
Entrez Gene IDs:
7040 (Human); 21803 (Mouse); 59086 (Rat); 397078 (Porcine); 100033900 (Equine)
Alternate Names:
CEDLAP; DPD1; latency-associated peptide; TGF beta; TGF beta1; TGFB; TGFB1; TGF-beta 1 protein; TGFbeta 1; TGF-beta 1; TGFbeta; TGF-beta-1; transforming growth factor beta-1; transforming growth factor, beta 1

Citation for Human Latent TGF-beta 1 ELISpot Development Module, 5 Plate

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.

1 Citation: Showing 1 - 1

  1. Human CD4(+) T cells recognize an epitope within alpha-fetoprotein sequence and develop into TGF-beta-producing CD4(+) T cells.
    Authors: Alisa A, Boswell S, Pathan AA, Ayaru L, Williams R, Behboudi S
    J. Immunol., 2008;180(7):5109-17.
    Species: Human
    Sample Types: Whole Cells

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