Hello Guys,
I am sharing one of my latest experience with you all which might be helpful for budding life science/biology researchers. I work on Colorectal cancer ...and carry out clinical studies too. Whoossshhhh...I know a lot to do. My interest is to understand the role of few proteins in contributing tumor progression. As a cancer biologists, we all know that KI-67 (antigen protein) is a nuclear protein which is associated with any cellular proliferation. Hence, used as Cancer cell marker protein. Now, here it is what happened. So I was following my usual protocol of immunofluorescence staining of animal tissue slide, and failed three times…(OMG). But this time, I thought of being lazy...and heated the slide for a longer duration for antigen retrieval step (and played some games during that time, I know that's wrong), used old antibody to tag. Next day, I came and did secondary antibody addition and went for imaging. I was very sure this time also it's going to be a waste of time as well as reagents (still I went on doing, because I had a very small ray of hope).
And guess what, thats was the best staining I ever did, saw beautiful images. I am sharing my happiness (see the image, sorry little blurred though). So now I was wondering which magic step it was? Starting from paraffin deparaffinization to dehydration in alcohol (= wanted so much after 3 failures), then rehydration. Later comes antigen retrieval step, where I became lazy (the magic step), so instead of heating 3 times...I did 4 times..so that I can relax more. After analysing everything step by step, and I found that it was the heating step. I was like…..WOW..even my protein wants to be HOT.
Even after being in research for 8 years, I still wonder which step or idea can work when and where!
P.S. I still don't know whether it was an idea or my laziness.
(If anyone interested I can share my protocol for Ki-67 IF staining)
Hope it was fun and informative too while reading this.
Love Ruby
Posted using Partiko Android
