* Antibody-coated beads supplied in compatible Bead Sets, sold separately (see below).
For performance data for the different analyte-specific beads, please see product datasheets for the compatible individual Beads Sets
NOTE: Due to a low level of cross-reactivity of the biotinylated anti-TGF-β1 antibody with TGF-β2, multiplexing TGF-β1 and TGF-β2 can result in falsely elevated values for TGF-β2 in some sample types. We found in human serum samples, falsely elevated values for TGF-β2 when multiplexing TGF-β1 and TGF-β2. Similar cross-reactivity was observed in competitor’s multiplex assays. For studies using human serum samples, TGF-β1 and TGF-β3, or TGF-β2 and TGF-β3 can be multiplexed; however, careful consideration should be given when deciding to evaluate TGF-β1 and TGF-β2 as a multiplex. Testing with samples from healthy donors found the impact of the cross-reactivity observed became insignificant with human milk, platelet-poor plasma, cell culture supernates and urine samples. It is suggested that each lab evaluate their samples to determine if multiplexing will be problematic (as TGF-β levels will vary with disease state and serum source for media).
Analyte | Catalog Number | Sensitivity (pg/mL) | High Standard Value* (pg/mL) | Microparticle Region | |
TGF-beta 1 | 11.1 | 33.3-24,300 | 34 | ||
TGF-beta 2 | 6.8 | 17-12,500 | 15 | ||
TGF-beta 3 | 14.3 | 68-49,850 | 22 | ||
*A standard curve must be generated each time an assay is run, utilizing values from the Standard Value Card included in the kit. |
Analyte | Cell Culture Supernate | Serum | Platelet-poor EDTA Plasma | Urine | Human Milk |
TGF-beta 1* | 1:5 | 1:15 | 1:15 | 1:15 | 1:15 |
TGF-beta 2 | 1:5 | 1:15 | 1:15 | 1:15 | 1:15 |
TGF-beta 3 | 1:5 | 1:15 | 1:15 | 1:15 | 1:15 |
*TGF-beta 1 is present platelet granules and is relesases upon platelet activation. Therefore, to measure circulating levels of TGF-beta 1, platelet-poor plasma should be collected for measurement. |
Magnetic multiplex kits are designed for use with the CCD Imager. Alternatively, kits can be used with the or Bio-Rad® Bio-Plex®, dual laser, flow-based sorting and detection platforms.
Analyte-specific antibodies are pre-coated onto color-coded magnetic microparticles. Microparticles, standards and samples are pipetted into wells and the immobilized antibodies bind the analytes of interest. After washing away any unbound substances, a biotinylated antibody cocktail specific to the analytes of interest is added to each well. Following a wash to remove any unbound biotinylated antibody, streptavidin-phycoerythrin conjugate (Streptavidin-PE), which binds to the biotinylated antibody, is added to each well. A final wash removes unbound Streptavidin-PE, the microparticles are resuspended in buffer and read using the Luminex MAGPIX Analyzer. A magnet in the analyzer captures and holds the superparamagnetic microparticles in a monolayer. Two spectrally distinct Light Emitting Diodes (LEDs) illuminate the beads. One LED identifies the analyte that is being detected and the second LED determines the magnitude of the PE-derived signal, which is in direct proportion to the amount of analyte bound. Each well is imaged with a CCD camera. Kits can also be used with Luminex 100/200 or a Bio-Rad Bio-Plex dual laser, flow-based systems.
Assays for the Luminex platform are offered as or . The Luminex High Performance Assays are fully validated panels with a focused selection of analytes. They are available in We Mix, You Mix, and Predetermined formats. The Luminex Assays allows for the maximum number of analytes in a multiplex and is supplied as a premixed kit.
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