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Hi Shane, I am measured decrees in NADH absorption in a recording uv/spectrophotometer for 3-5 min, at 340nm. On Wed, Aug 5, 2015 at 11:03 PM, Shane Caldwell <[log in to unmask]> wrote: > Hi Rohit, > > How does your assay work? Iodide can interfere with spectroscopic methods. > It's also more reactive than F, Cl, or Br, and can participate in redox > chemistry. Without additional information, I'm afraid it's really difficult > for anyone to make any concrete recommendations... > > Shane Caldwell > McGill University > > > On Wed, Aug 5, 2015 at 12:52 PM, rohit kumar <[log in to unmask]> wrote: > >> Dear All, >> >> I am trying to check the affect of different halide on our enzyme. The >> reaction assay have >> 50 mM TAPS, pH 8.5, 0.2 mM NADH, 1.0 mM DTT, 1.0 mM EDTA, 5.0 mM >> substarte1, excess amount of PGDH, 5.0 mM substrate 2, and 4 µg of >> enzymes (enzyme of interast). I am using 200 mM of halids (NaF, NaCl, NaBr >> and NaI) in separate reactions. >> >> In prsence of 200 mM NaF, NaCl, and NaBr the enzyme activity is >> not affected. But in case of NaI > 50% activity is decreased. >> >> our enzyme does not have any halide binding site. >> >> Can any one just tell me why NaI give such affect? >> Is it because of bigger size of I (iodide ion)? or something else.... >> >> >> Please suggest >> >> >> >> -- >> WITH REGARDS >> >> Rohit Kumar Singh >> >> Research Scholar >> > > -- WITH REGARDS Rohit Kumar Singh Lab. no. 430, P.I. Dr. S. Gourinath, School of Life Sciences, Jawaharlal Nehru University New Delhi -110067