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14 Cards in this Set
- Front
- Back
when is the specific activity of a particular enzymatic activity of particular importance?
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when proteins are being purified or different sample are being compared.
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what do the most utilized methods to assay total protein rely on
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the reduction of copper in the presence of a chromogenic reagent
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_____ is the most widely used method for accurate determination of protein concentration
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lowry's method of protein estimation
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lowry's method is a combination of what two reactions?
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biuret and folin ciocalteau
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what is the first step of the reaction
and in the second step? what does this reaction produce and what does it predominantly depend on |
protein binds to copper in alkaline medium and produces cu++
cu++ catalyses oxidation of aromatic amino acid by reducing phosphomolybdotungstate to heteropolymolybdanum blue strong blue color tyrosine and trytophan content of protein to a lesser extent cysteine |
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why do we add BCA reagent after lysing cells?
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to provide copper ion which is going to adhere and turn blue
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copper absorbs light at __nm
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562
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transmittance is how much is ____
the spectrophotometer reads out ____ how do you find OD |
emitted
absorbance 1/log(A) |
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_____ are going to absorb light from spectrophotometer
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buffer reacted with protein
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what are the units for conc of protein
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micro grams/mL
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standard is a known quantity used as a control
how can we use this |
based of known we can predict unknown samples
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the higher the OD the ____ the proteins
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higher
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what was the blank 50ml buffer used for
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to calibrate the spectrophotometer
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the concentration of total protein from the cells contain what kind of proteins
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membrane nuclear and cytosolic proteins
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