How to Make a Lineweaver Burk Plot in Excel (with Easy Steps)

In biochemistry, the Michaelis-Menten Equation of enzyme kinetics results in a Lineweaver-Burk Plot, also known as a Double Reciprocal Plot. Here’s an example of one such chart.

How to Make a Lineweaver Burk Plot in Excel

Lineweaver Burk Plot and Its Components

What Is a Lineweaver-Burk Plot?

A Lineweaver-Burk Plot is the graphical representation of the Michaelis-Menten Equation. The plot is used to determine different types of inhibitions.

Components of Lineweaver Burk Plot

Substrate Concentration

Substrate Concentration, S. X-axis of the Lineweaver Burk plot that is reciprocal of substrate concentration, [1/S].

Initial Velocity

Initial Velocity during an enzyme-inhibited reaction, V or Vo. Y-axis of the Lineweaver Burk plot that is reciprocal of velocity, [1/Vo].

Maximum Velocity

Maximum Velocity of the enzyme-inhibited reaction, Vmax. Y-axis interception of the plot is reciprocal of maximum velocity, [1/Vmax]. 

Michaelis Constant

Michaelis Constant, Km is the measurement of enzyme affinity. X-axis interception of the plot is reciprocal of Michaelis Constant, [-1/Km].

How to Make a Lineweaver-Burk Plot in Excel: Step-by-Step Procedure

Making a Lineweaver-Burk plot requires data on Substrate Concentration (S) and Initial Velocity (Vo).

Step 1 – Setting Up the Data

  • Compile the raw substrate concentration and initial velocity data as depicted in the picture below.

Data for making a Lineweaver Burk plot

  • Find the reciprocals of both points (i.e., S and Vo) as shown.

Read More: How to Plot Semi Log Graph in Excel

Step 2 – Inserting a Scatter Plot

  • Highlight the reciprocals, leaving the first non-value entries, and then go to Insert Scatter (inside Charts section).
  • Click Scatter.

  • Excel inserts a Scatter Plot as shown below.

Read More: How to Plot Time Series Frequency in Excel

Step 3 – Modifying the Scatter Plot to Make a Lineweaver-Burk Plot in Excel

Stretching the Trendline Backward results in a Lineweaver-Burk plot.

  • Click on a point in the plot, then right-click on it. The Context Menu appears.
  • Select Add Trendline.

  • Excel brings up the Format Trendline side window.
  • Select Linear (Under Trendline Options)
  • Enter the Backward value 0.07 or any suitable value.
  • Check Display Equation on Chart.

Modifying the Scatter Plot as a Lineweaver Burk Plot in Excel

  • Choose a plot design. Here’s an example.

Read More: How to Make a Time Series Graph in Excel

⧭ Notes: The Lineweaver-Burk Plot may have a different depiction depending on the intensity of its components. Therefore, the following image displays multiple Lineweaver-Burk plots distinguishing between different inhibition reactions.

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Maruf Islam
Maruf Islam

MARUF ISLAM is an excellent marine engineer who loves working with Excel and diving into VBA programming. For him, programming is like a superhero tool that saves time when dealing with data, files, and the internet. His skills go beyond the basics, including ABACUS, AutoCAD, Rhinoceros, Maxsurf, and Hydromax. He got his B.Sc in Naval Architecture & Marine Engineering from BUET, and now he's switched gears, working as a content developer. In this role, he creates techy content... Read Full Bio

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