How do you analyze a western blot in ImageJ?

1. Open the western blot image in ImageJ. 2. Make sure that the image is in 8-bit mode: go to Image>Type>8-bit….Data analysis

1. Place your data in a spreadsheet.
2. In a new column next to the Percent column, divide the Percent value for each sample by the Percent value for the standard (the 1st peak in this case, 26.666).

How do you analyze a western blot band?

The 4 Important Steps for Western Blot Quantification

1. Find the Linear Range. For quantitate analysis of an image you must ensure your image was captured in a manner sensitive enough to detect change, in what we call the “linear range”.
2. Subtract Background.
3. Normalize.
4. Graphs and Stats.

How do you quantify western blot bands with ImageJ?

You can quantify by the following steps:

1. Open western image in Image J, select Rectangular Selections tool from the ImageJ toolbar and select first western band.
2. Press Ctrl +1 and drag the same rectangle selection to the next band and press Ctrl + 2.

How do you do a densitometry in western blot?

Densitometry Western Blot

1. Open Western scan in Image.
2. Click the ‘rectangular selection’ (under file menu) and select a box around a band of interest.
3. Once you have boxed an area go to the ‘analyze’ drop menu.
4. Use the arrow curser to move your box along to the next band of interest, and repeat the ‘measure’.

How to do western blot analysis in ImageJ?

Figure 1. A fabricated western blot image opened in ImageJ. The information along the top of the image indicates that the image is currently in 8-bit mode, using an inverting LUT (look-up table). The inverting LUT ensures that dark bands will be recorded as higher density values. 3. Choose the Rectangular Selections tool from the ImageJ toolbar.

Which is the best software to analyze western blots?

However, the free Image Studio Lite is more than sufficient for basic analysis of western blots, as I’ll show below. Obviously you need to start by downloading and installing Image Studio Lite.

How to convert a western blot to grayscale?

The simplest method to convert to grayscale is to go to Image>Type>8-bit. Your image should look like Figure 1. Figure 1. A fabricated western blot image opened in ImageJ. The information along the top of the image indicates that the image is currently in 8-bit mode, using an inverting LUT (look-up table).

What are the peaks in a western blot image?

In the example western blot image, the peaks in the profile plot (Figure 4) correspond to the dark bands in the original image (Figure 3). Because there were four lanes selected, there are four sections in the profile plot. Higher peaks represent darker bands. Wider peaks represent bands that cover a wider size range on the original gel. 10.