BIOCHEMISTRY

Determining the isoelectric point (pI) of an amino acid

Introduction: Isoelectric point (pI) is the pH at which the net electric charge on a molecule/protein is zero. At its pI, the molecule/protein is electrically neutral that the molecule at its pI exists as a zwitterion with an equal number of positive charges (from protonated amino groups) and negative charges (from deprotonated carboxyl groups). At […]

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Estimation of Protein by Biuret Method: Principle, Procedure, Observation Table, Result

Aim: To estimate the concentration of protein using Biuret method. Principle: The is a colorimetric estimation technique that is specific for proteins and peptides used for estimation of protein. The coppers salt reacts with the two more peptide bonds under alkaline condition result in purple complex. The intensity of the formed color complex is proportional

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Estimation of Protein by Bradford Method:Principle, Procedure, Observation Table, Result

Aim: To estimate the concentration of protein using Bradford Method. Principle: The Bradford protein assay is a commonly used method for estimating the concentration of proteins in a sample. Generally, it is based on the binding of Coomassie Brilliant Blue (CBB)  dye to proteins, resulting in a shift its  maximum absorbance maximum from 465 nm

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Separation of Plant Pigments by Thin layer chromatography (TLC)

Aim: To separate green leaf pigments (Spinach leaves) by Thin Layer chromatography (TLC) method. Introduction: It is a type of planar chromatography, in which the stationary phase is a thin layer of adsorbent particles attached to a solid plate. A small amount of sample/analyte is applied (spotted) near the bottom of the plate (an imaginary

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Quantitative estimation of reducing sugar (Glucose) by DNS method.

Introduction: The DNS (3,5-dinitrosalicylic acid) method is commonly used for the quantitative estimation of reducing sugars, including glucose. This method is based on the reaction of reducing sugars with DNS reagent, which results in the formation of a coloured compound that can be measured spectrophotometrically. Principle: The principle behind the DNS (3,5-dinitrosalicylic acid) method for

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Enzyme Immobilization/ Immobilized enzyme: Introduction, Methods, Factors, Kinetics,Applications

Introduction: Benefits of immobilized enzyme: Fig: Application of immobilised enzyme Properties of carrier matrix for ideal immobilization: Biocompatibility: The carrier matrix needs to be biocompatible in order to maintain the activity, stability, and conformation of the specific enzyme. It shouldn’t cause the enzyme to become denaturized or inactive. Chemical Stability: The matrix should be chemically

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