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Isoelectric Point Calculator

Isoelectric Point Calculator

Calculate the pH at which molecules have zero net electrical charge

Calculate Isoelectric Point

Acid dissociation constant (-log Ka)

Base dissociation constant (-log Kb)

Formula Used

pI = (pKa + pKb) / 2

Calculation: (0 + 0) / 2 = 0.00

Results

0.000
Isoelectric Point (pI)
Strongly Acidic
Classification
Negative at physiological pH
Charge at pH 7.4

Description: Very low isoelectric point indicating strong acidic character

Solubility: Poor solubility at neutral pH

Example Calculation

Glycine Amino Acid

Given: pKa = 2.34, pKb = 9.60

Formula: pI = (pKa + pKb) / 2

Calculation: pI = (2.34 + 9.60) / 2 = 11.94 / 2

Result: pI = 5.97

Step-by-Step Calculation

1. Add the pKa and pKb values: 2.34 + 9.60 = 11.94

2. Divide the sum by 2: 11.94 / 2 = 5.97

3. The isoelectric point is 5.97

4. At pH 5.97, glycine has zero net charge

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pH Classification Guide

pI < 3

Strongly acidic

Poor solubility at neutral pH

3 ≤ pI < 5

Acidic

Negative charge at pH 7.4

5 ≤ pI < 8

Neutral

Near neutral at pH 7.4

8 ≤ pI < 11

Basic

Positive charge at pH 7.4

pI ≥ 11

Strongly basic

Poor solubility at neutral pH

Applications

Protein Purification
Ion-exchange chromatography
Electrophoresis
Protein separation by charge
Drug Formulation
Solubility optimization
Crystallization
Protein structure studies

Understanding Isoelectric Point

What is Isoelectric Point?

The isoelectric point (pI) is the pH value at which a molecule has zero net electrical charge. At this point, the molecule is electrically neutral, with equal numbers of positive and negative charges.

Key Concepts

  • Zero Net Charge: Equal positive and negative charges
  • pH Dependent: Charge state changes with pH
  • Solubility Minimum: Lowest solubility at pI
  • Zwitterion: Molecule with both + and - charges

Calculation Formula

pI = (pKa + pKb) / 2

  • pI: Isoelectric point
  • pKa: Acid dissociation constant (-log Ka)
  • pKb: Base dissociation constant (-log Kb)

Note: For amino acids with ionizable side chains, the calculation involves additional pKa values and becomes more complex.

Applications and Importance

Protein Purification

Ion-exchange chromatography separates proteins based on their charge at different pH values relative to their pI.

Electrophoresis

Proteins migrate toward opposite charges in electric fields. At pI, migration stops.

Drug Development

Understanding pI helps optimize drug solubility and bioavailability at physiological pH.

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