Question

In: Biology

Make a comparative chart on the main methods of protein quantification. Method Reagents Equipment characteristics Bradford...

Make a comparative chart on the main methods of protein quantification.

Method
Reagents
Equipment
characteristics
Bradford
Biuret
Lowry

BCA (bicinconinic acid)

Solutions

Expert Solution

Method

Regents

Equipments

Characteristics

Bradford

Coomassie Brilliant Blur G-250

Preparation:

  1. Dissolve 100mg of G-250 in 50ml (95%) ethanol + 100ml (85%) phosphoric acid. After complete dissolution dilute it to 1L with water and then filter

Light spectrophotometer (that can detect 595nm wavelength), Cuvettes

It is a very fast, sensitive and accurate method for the estimation of protein

Biuret

Na-K-tartrate, CuSo4.5H2O, KI,

Preparation:

Take 9g (Na-K-tartrate) + 3g (CuSo4.5H2O) +5g (KI) in 500ml 0.2 M NaOH

Light spectrophotometer (that can detect 550nm wavelength), Cuvettes

It is time taking and low sensitive method

Lowry

Na-K-tartrate, CuSo4.5H2O, Folin reagent, NaOH, Na2Co3

Preparation:

  1. Reagent-A: 2g (Na-K-tartrate) + 100g (Na2Co3) in 1L 1N HaoH
  2. Reagent-B: 2g (Na-K-tartrate) +1g (CuSo4.5H2O) in 0.1N NaoH
  3. Dilute 1: 15 Folin-Ciocalteau reagent

Light spectrophotometer (that can detect 650nm wavelength), Cuvettes

It is sensitive and time taking methods but used in which various interfering components along with proteins present in the analysing solution, This is a two-step process so needed more time

BCA (Bicinconinic acid)

BCA, Na-K-tartrate, CuSo4.5H2O, NaOH, Na2Co3

Preparation:

  1. Reagent-A: 1g (BCA) + 2g (Na2Co3) + 0.16g (Na-K-tartrate) + 0.4g (NaOH) + 0.95g (Na2Co3) in 100mL, pH should be 11-12
  2. Reagent-B: 0.4g (CuSo4.5H2O) 10ml H2O

Light spectrophotometer (that can detect 562nm wavelength), Cuvettes

As similar to the Lowry method. It is a single-step process so less time and technical error possible


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