To dampen immune responses, the anti-inflammatory cytokine TGF-beta can be produced by macrophages. However few good detection methods are present or reproducible to detect this cytokine. Anyone used a good antibody to detect the secretion of this cytokine (or a publication)?
The lack of a decently performing assay may be due to the fact that TGF-beta is secreted in a "wrapping" (LTBP and LAP). This package leaves TGF-beta inactive and probably difficult to detect by ELISA directed at TGF-beta. The critical point for TGF-beta activity is not necessarily secretion, but subsequent activation by cleaving off these two polypeptides. Unless you produce an assay for both the "wrapped" TGF-beta and the active form, you will have undetected TGF-beta that may bias your experiment
True, the obtain an active form of TGF beta one could acidify the supernatant, however, it remains a tricky method. I was hoping an alternative would have been on the market by now.
I recommed R&D system antibodies (anti TGF β1 - Cat No AB 101 NA) and Elisa Kits.
For alternative activation methods and elisa option see attached files.
I would recommend the Meso Scale Doscovery kit. Always very good performance for many different matrices.
I will suggest both R&D and BD Sciences ELISA kits to detect the cytokine.
Go with any system which uses Clone 1D11 - originally developed by Jim Dasch at Collagen Corporation. It recognizes and neutralizes all three isoforms across most mammalian species. Acidification of the sample is required to dissociate LAP from mature TGFb
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