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enzymepurifier

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    Protein Biochemistry

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  1. Sorry, for the confusion - I am still a little confused I think... If we are hypothetically working with an ion exchange stationary phase (say SP resin), is the analyte:SP resin affinity in anyway correlated to to when it would elute off the column in a 0-1M NaCl gradient? i.e. If the analyte elutes early in the gradient would that indicate weaker affinity than if it eluted late in the gradient? I think you already answered this actually --- but, if this is true wouldn't binding capacity of a stationary phase be based on the strength of affinity to that specific analyte?
  2. Is there any relationship between total resin binding capacity for a "target" molecule and when in a gradient elution that "target" molecule elutes off? In other words, if a molecule elutes off earlier in a gradient, is that an indication of lower binding capacity relative to a different target that would elute off higher in a gradient with the same conditions?
  3. First, does your diluted measurement fall within the high and the low points of the standard curve that you generated? If so, you most certainly have to take into account the dilution that you made initially. Option "B" is correct in that you will have to multiply the output value by your dilution factor of 10. e.g. I have a standard curve that measures 0.000 OD to 1.000 OD that generated a working concentration range of 0 mg/ml to 10 mg/ml. If I made a 10x dilution of my sample and it read 0.500 OD, the resulting concentration would be 50 mg/ml.
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