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im interested to know what your looking at since ive been nagging my supervisor for some time now to get me TGF-B3 reagents.
How sure do you need to be its active TGF-B3? you can get a very reliable TGF-B3 ELISA kit from R and D systems, its pricey, but will attach to only TGF-B3. Whether the latency associated peptide (LAP) will interfere with this i dont know. Maybe run a weswtern blot, and you'll get different bands for your active TGF-B3 and TGF-B3/LAP complex, although the TGF-B3 will be ahrd to distinguish from the other isotypes as theyre very close in molecular weight. as far as i know, when i was doing a TGF-B1 ELISA, the acid step preparing my samples probably activated my latent TGF-B1, so it wasnt a true indication of how much ACTIVE TGF-B was in my sample at that time, more an indication of the quantity that had been made but was in different states waiting activation.
hope that helps. basically i'd say either a very expensive ELISA kit (around £450 for a 96 well plate kit), or a western blot (you can get antibodies for around £200 per 100ul).
is there a way of checking for active TGF-B3 based on binding to/activation of a TGF-BR?
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