We've been using 'instant' kit by R&D for mouse TGFb which is also validated for rat. For procedure - follow the datasheet. It's good to check also latent TGFb level ;)

http://www.rndsystems.com/Products/MB100B

See articles Renee van Dorp Elisa and microwave techniques.

We at our lab use BD Biosciences kit for quantification of TGF-beta. It works quite well. The other options include kit from e-biosciences. The procedure is quite simple. Just coat primary antibody in 96 well plate supplied with the kit in the coating buffer overnight  at 4 degree, followed by 3 washes with PBSt. Then add standard and samples to the wells, incubate for an hour at 37 degree and wash with PBST. Now add HRP conjugated secondary antibody, incubate for an hour at 37 degree, followed by washings. Finally add substrate and wait for 10-15 minutes for color to develop. Stop the reaction with 2N H2SO4 and read plate using  a microplate reader.

We used to quantify rat TGF-beta1 with the Human DuoSet kit from R&D System. It works quite well, is cheep and easy to use.

For note, we also try a kit from Promega, but it failed to detect our sample.

I am also interested in assaying for active TGFB1 in whole brain homogenates from mice. I have two questions, and would appreciate your help:

1) How did you homogenize your brain tissue for the TGFB1 ELISA? If you have a buffer recipe you found appropriate, I would really value if you shared it with me.

2)When using commercial ELISA kits for TGFB1, such as the kits from R&D that are not typically validated for tissue homogenates, did you follow the sample activation steps for cell culture supernatant or the steps for plasma/serum? Similarly, what samples diluent did you end up using? As this kit has two sample diluent buffers, one if using cell culture supernatants and one if using plasma/serum samples, none if using tissue homogenates.

Manual for R&D kit:https://resources.rndsystems.com/pdfs/datasheets/mb100b.pdf

Thanks,

Rania

Rania Faidi hi! did you manage to find answers for your questions? I am struggling to detect TGFb1 (both active & latent) in mouse brain lysates (we chop the brain and take an aliquote for lysis and the rest for flow cytometry). please let me know - any feedback would be helpful! thanks, Sal