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.

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 V_o. Y-axis of the Lineweaver Burk plot that is reciprocal of velocity, [1/V_o].

Maximum Velocity

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

Michaelis Constant

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

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 (V_o).

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 V_o) as shown.

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.

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.

⧭ 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.