Lidstrom:Choosing a protein concentration quantification method

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== Major methods available ==
== Major methods available ==
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* Spectrophotometer: use a formula based on A260 and A230 (260 and 230 nm absorbances)
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=== Spectrophotometer ===
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** fastest/easiest but may have most bias (??)
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* Absorbance at 280 nm gives a quick approximate estimation of protein in non-turbid solutions, which is often all that is required. [http://www.amazon.com/Enzyme-Assays-Practical-Approach-Series/dp/0199631425/ref=sr_1_1?ie=UTF8&qid=1383921457&sr=8-1&keywords=0199631425 ISBN 0-19-963142-5]
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** Ceci/Amanda/Frances chose BCA instead.  
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** Advantage" non-destructive.
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* Pierce brand BCA assay
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=== Others ===
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* Since the 1950s when Lowry et al. (8) modified the method originally devised by Wu (9) and Folin and Coicalteu (10), the Lowry method has become the most widely used for protein estimation.  However, during the 1980s  dye-binding assays based on Coomassie Blue (11,12) have become more popular because of their simplicity and because of certain shortcomings in the Lowry method.  One of the oldest procedures, the Biuret test (13), is still used; it is quite satisfactory apart from the disadvantage of being relatively insensitive. 
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=== Who uses what as of 2013 ===
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* Mila uses the spectrophotometer.  She thinks hers is based on A260 and A230 (260 and 230 nm absorbances)
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* Ceci/Amanda/Frances chose BCA, using a pierce brand kit.
== Compatibility with your lysis solution ==
== Compatibility with your lysis solution ==

Revision as of 10:34, 4 December 2013

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Contents

Major methods available

Spectrophotometer

  • Absorbance at 280 nm gives a quick approximate estimation of protein in non-turbid solutions, which is often all that is required. ISBN 0-19-963142-5
    • Advantage" non-destructive.

Others

  • Since the 1950s when Lowry et al. (8) modified the method originally devised by Wu (9) and Folin and Coicalteu (10), the Lowry method has become the most widely used for protein estimation. However, during the 1980s dye-binding assays based on Coomassie Blue (11,12) have become more popular because of their simplicity and because of certain shortcomings in the Lowry method. One of the oldest procedures, the Biuret test (13), is still used; it is quite satisfactory apart from the disadvantage of being relatively insensitive.

Who uses what as of 2013

  • Mila uses the spectrophotometer. She thinks hers is based on A260 and A230 (260 and 230 nm absorbances)
  • Ceci/Amanda/Frances chose BCA, using a pierce brand kit.

Compatibility with your lysis solution

protein assay compatibility table from Pierce
protein assay compatibility table from Pierce

Resources

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