Hi all,
I'm working on reading the amount of HSA adsorbed onto a hydrogel with SPR, quantitatively and I need to confirm the results using at least two other methods.
What would be a good suggestion to do so? I saw that the tagging process for western blotting is long and hard needing a lot of techniques, ELISA was easier to go, but I was thinking of maybe spectrophotometry and florescent tagging the HSA itself, instead of buying its anti-antibody. However The protocols I found were radioactive and not so much on flouroscent tagging a protein, as if it has gone out of date.
Please let me know of any good protocol or reviews if you may.
Thank you.
I definitely recommend fluorescent tagging. For HSA, any generic kit you purchase (e.g., an amine-reactive fluorophore) should work perfectly well for spectrophotometric quantitation. If you have access to an HPLC, you could pretty easily quantify HSA content, and I'm sure there are publications out there that have developed run conditions, so you wouldn't need to develop one from scratch.
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