Revolutionize Your Western Blot Analysis: Learn How to Perform Densitometry with ImageJ

#Westernblot #eletrophoresis #analysis #densitometry #imagej
How to analyse Western Blot bands densitometry with the most simple ImageJ method.

[@jamersoncarvalho4330]

Que orgulho que você é Br! Vídeos maravilhsos, obrigado por compartilhar

[@bioinfotips]

Obrigado Jamerson mas na verdade sou Português PT. Fico contente pelos vídeos estarem a servir o seu propósito. Se tiveres algum assunto que queiras ver abordado não hesites em comentar :)

[@jamersoncarvalho4330]

@@bioinfotips ah, continua maravilhoso rs estou explorando ainda os demais vídeos, se tiver algo nesse sentindo volto a comentar! Obrigado😁

[@nicksmith7750]

thank u for this video king

[@bioinfotips]

Glad it was helpful to you nick!

[@chatty_homebody]

Thanks for making this video! I was wondering how to do densitometry if I don't have the control to calculate the adjusted ratio. So I western-blot for proteins of interest (including GAPDH as a loading control) from different cell lines without any treatments. Do I just stick with the POI/loading control densitometry calculation?

[@bioinfotips]

Thank you for the comment! In that case, I think the best approach is probably to normalize it to one the different cell lines. Alternatively, you can just present the ratios without normalizing it. One other option would be an ELISA to have the absolute quantification and not a semi-quantitative one.

[@FindMeInsanity]

thanks for the video! however i dont understand what shall i do if the curve doesnt touch the x-axis. may you please suggest what should be done in such scenario ?

[@bioinfotips]

You welcome. Thank you for your question. However, I don't see a scenario where that happens... Can you send a print screen?

[@Yhusi]

Thank you very much
May I know how to combine all data from different membranes?

[@bioinfotips]

If it's just the same layout in the membrane like: Control, treatment1, treatment2... You can put all the densitometries in Excel or graphpad and make a bar chart with the mean and standard deviation. Let me know if this answered your question.

[@jessicacolon-gonzalez8503]

thanks!

[@bioinfotips]

You welcome!

[@tubaoz8903]

Thanks for wonderful video ! but ı dont understand why you write 1000/2000/3000 etc.

[@bioinfotips]

Thank you so much for your feedback. The 1000, 2000 are an example of possible values for a target protein. Since the example I analyzed in the video was a beta actin blot, I made up those values just to give an example on how to analyse it. Let me know if this was clear.

[@Ahmad90n]

although useful , but ...
1. we know it is youtube and we can pause the video, but YOU DON'T HAVE TO BE THIS FAST!!!! ok you are great we know
2. why the hell excel sheet is this small and unclear, I had to repeat same second many times and I barely get what you did!!!
3. your voice is sleepy and not clear

[@bioinfotips]

Thank you for your feedback. This is one of the first videos I did. I tried to improve in each and every one of them ever since.