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How does aggregation influence absorbtion at 280nm?


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#1 nenette

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Posted 15 February 2010 - 08:44 AM

Hi!

If a protein aggregates, does it absorb the same way at 280 nm or does it drop/increase in absorption?
Can one say anything generally here... ?

#2 paramyosin

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Posted 23 February 2010 - 01:19 AM

When we purify a new protein always make a meassurement at 230 nm due to (theoretically) the ratio 230/280 gives you the agreggation %. Nearly always, some % of your 280 absorbance is light scattering due to the presence of aggregates, it is not strange to obtain a 10-25% of aggregates in a purification procedure. Sometimes, if you change the buffer or make use of several specific aditives (detergents, DTT, caothopic agents) that improve the solubility, the % of aggregates diminish and your absorbance at 280 nm goes down also.
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