I wish I had this explanation a long time ago! Thank you for sharing this knowledge. Doing experiments is cool, but analyzing data is always more tricky!

Thank you! It was very clear and systematic-easy to understand and follow.

Final ratio was rather Protein r/ actin r I think. Good video! thank you.

Sorry, just need a clarification - do you divide Actin Ratio by Protein Ratio or vice versa? You wrote down Actin R/Protein R but do otherwise. Thanks!

Excellent job!!! Very clear and consize! Many thanks!

Excellent explanation sir . Thank you very much for your video

Many thanks, once again. Please, how do I obtain the relative intensity (Protein expression) for samples if the control does not have any protein expression?

Thank you so much for making it easy to understand.

thank you for saving my life. awesome.

Thank you so much for sharing your knowledge with us. I really appreciate your work. Sir, i have one query. You have taken 1 experimental control for explaining the analysis process. Sir, i have done western blot on clinical samples and i have taken 3 control samples and 3 patient samples for my western blot process. So the thing is how will i take the ratio for those 3 controls?

Excellent explanation. Thanks a lot.

Thank you for the clear explanation! I had one question, what if my CTRL value is 0, how do I now calculate the ratio's and perform the normalisation?

Thanx for responding my question! I subscrbed, liked + shared your video

Excellent explanation sir. This is what I needed

Thank you! Helped a lot

Hi thanks for the video. If my control protein and protein of interest are in seperate images will the contrast settings affect the result?

I did a western blot with n=4 for both control and treatment to observe the expression of a protein of interest and then normalise against housekeeping gene .. how do I plot the data in this case? Do I calculate the ratios for each n individually? how do I carry out a paired student t-test with ratios o does it need to be with the means of the area under the curve calculated with image j? I would appreciate your help. thanks a lot!!

Thank you for such a beautifully explained video.

I can't drag the rectangles just like you did. I tried so many times already

Very good explanation. Thank you!

Many thanks! I observed that some faint bands were entirely eliminated due to the adjustments of brightness/contract; how can you account for the bands. What are they, please?

At 4:22 you said there was no need for you to adjust the lanes. What I get are lanes that "don't touch the zero" so two bands have the same curve. How do I separate them?

Thank you for the video, how do you tag the Y axis in the plot?. Is it ok to label it like: Protein/actin (relative to control)?.

Life saver. Thank you so much sir. Subscribed.

Very helpful!!

If we quantify western blot like this, our Y-axis should label relative expression, right?

hi so if i do it like this, i do not need to subtract the background?

What if I only am blotting for 1 protein? How would I analyze this data? I did a WB on A549 cells, probing for beta-actin.

I don't think it's a good thing to play on intensities like that with contrast

So informative...thank you

Thank you!

Thank you

any reffrence to this technique, pls provide as early as possible