How do you quantify a western blot band?
Step 1: Determine the background-subtracted densities of your protein of interest (PI) and the normalizing control (NC). Step 2: Identify the NC that has the highest density value. Step 3: Divide all the NC values by the highest NC density value to get a relative NC value.
Can western blot be quantitative?
Western blot is a reliable quantitative method only if sample properties and integrity, antibody specificity to the target protein, and loading protocols are considered. With careful attention to details, you can avoid common mistakes and avoid misinterpreting Western blot data.
How do you quantify western blot bands in ImageJ?
You can quantify by the following steps:
- Open western image in Image J, select Rectangular Selections tool from the ImageJ toolbar and select first western band.
- Press Ctrl +1 and drag the same rectangle selection to the next band and press Ctrl + 2.
How do you present data in western blot?
When presenting a western blot in a Starr lab meeting or presentation, include the following information: Title: Date, protein(s) and cell lysates including conditions being analyzed. Subtitle: Your initials, and date where details can be found in your lab book (see Lab Book Details).
Why is western blot semi quantitative?
Quantification. It is very important to be aware that the data produced with a western blot is typically considered to be semi-quantitative. This is because it provides a relative comparison of protein levels, but not an absolute measure of quantity.
What is quantitative western blot?
What is a Quantitative Western Blot? A quantitative Western blot makes relative comparisons between different treatments possible. The goal of a quantitative Western is to accurately measure changes in protein expression.
What is band intensity?
The different vibrations of the different functional groups in the molecule give rise to bands of differing intensity. The absorptivity is an absolute measure of infrared absorbance intensity for a specific molecule at a specific wavenumber.
