Hello again! My previous post discussed how I finally had a specific antibody to EZH1 in a cell line (HEK293). As a follow up, I wanted to determine if we could detect EZH1 expression in patient cells. I’m happy to report I was able to detect the EZH1 protein in patient cells (Figure 1). Experimental details can be found on Zenodo. We can now use this antibody to determine EZH1 expression in other patient cells.
Figure 1: Western blot for EZH1 (green) and actin (red) with ~50ug protein loaded in HEK293 cells (293) and patient cells. A band of expected size (~85kDa) is seen in all samples except 150860 that we did not expect EZH1 expression.
Something else that I wanted to share was members of SGC Toronto (including other open notebookers Rachel, Mandeep and Dave) recently participated in the Leukemia and Lymphoma Society of Canada’s (LLSC) Light the Night Walk. The walk is the LLSC’s annual fundraising event to raise money to support those impacted by blood cancers, as well as funding for leukemia and lymphoma research. LLSC is a great organization that funds my research at the SGC, including the work posted on my open notebook. Through the generosity of our supporters, team SGC was able to raise over $2000 for LLSC! It was a truly memorable experience seeing all the lanterns lighting up the night in support of fighting blood cancers and I definitely won’t forget the touching stories we heard from survivors, patients and their loved ones battling leukemia and lymphoma.
I would *strongly* suggest you engineer an EZH1 knock-out line as a negative control and do not use mobility on a gel as a signal of a positive antibody. In our antibody validation projects we have identified quite a few antibodies that recognize a band of the right size…only to see the same band in the knock-out lines. It’s really worth the effort
Genna Luciani, An interesting article Dec. 21, 2018 about pediatric leukemia in Northwestern News @ https://news.northwestern.edu/stories/2018/december/pediatric-leukemia-drug/ Enjoy 🙂