2. Provide an explanation for the following steps that you...

2. Provide an explanation for the following steps that you did: Using Bradford reagent? measuring the absorbance at 595 instead of 280? 3. You measured the absorbance at 280 nm and 595 nm, Which possible molecules or function groups or complexes that may be responsible for the absorbance at these wavelengths?. 4. According to the standard curve, what is the relationship between the absorbance and the concentration ?. 5. Are the concentrations of your unknowns fall within the concentration range of your standard curve ? if not what you have to do?. 6. What is the advantage and disadvantage of the method that you used to quantify the protein concentration ? compare this method with other methods for protein quantification ?.

Transcript

00:01 Hello students, so the first part of the question is to provide an explanation for and the steps that you will do by using bradford reagent.

00:10 So here the bradford reagent is the assay, which is a calorimetric method and it is used to determine to determine the concentration concentration of the protein concentration of the proteins.

00:36 So here this reagent consists of the comasi brilliant blue g250 dye and this binds to the protein causing the shift in the dye's absorbance.

00:50 So this causes the shift the shift in the dye in the dye's absorbance absorbance maximum from 465 nanometer to 595 nanometer.

01:18 This shift in the absorbance can be measured by the spectrophotometer.

01:24 Now here this is used and the concentration and the protein concentration can be determined by comparing the absorbance to the standard curve.

01:34 Now measuring the absorbance at 595 nanometer.

01:40 So here the absorbance at 280 nanometer is typically used to measure the protein concentration of based on the absorbance of the aromatic amino acids.

01:52 So now let's move towards the third part of the question.

01:56 So here in this question, the question is why the the question is which possible molecules or the functional group of complexes can be responsible for the absorbance at at 280 nanometer and 595 nanometer.

02:15 So here the answer is so the molecules or the functional groups, which is responsible to absorb at this spectrum at this wavelength is primarily due to the presence of this happens due to the presence of presence of the aromatic aromatic amino acids such as tryptophan, tryptophan, tyrosine, etc.

02:59 And the peptide bonds which consist of the aromatic ring...

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