Follow up to finding an antibody to detect EZH1 protein expression…the search continues

Following up on my previous post, I tested two antibodies I was planning on trying from Abcam and Novus that were used in previous publications. This time I tested directly in patient sample lysates and either saw no band, or many nonspecific bands (Figure 1). I have since reached out to the Orlando lab who Read More …

Optimizing methodology for the detection of H3K27me3 levels using flow cytometry

Hello again! In previous posts I talked about how we were using a co-culture system to grow our leukemic cells. Although this is a great way to maintain cells, it can complicate some analyses where we cannot distinguish mouse (stroma) from human (AML) cells. We wanted to determine H3K27me3 levels (the mark that EZH2 is Read More …

Determining the plasticity of CD34 expression

Previously, we established that OCI-AML-20 requires stroma to maintain CD34 expression (https://zenodo.org/record/1187083#.Wr5-1C7wbIV). In this experiment, we wanted to see whether those cells grown in suspension are able to regain CD34 expression when cells are reintroduced to stroma. We find that OCI-AML-20 regains CD34 expression when put back on to stroma (either MS5 or OP9). Full Read More …

Determining CD34 expression in the presence and absence of stroma

We are currently working on a manuscript to characterize one of the cell lines that we established from growing patient samples. I previously discussed how we established OCI-AML-20 and I’m currently working on some experiments to complete the manuscript. One of those experiments is determining whether the CD34 expression seen in OCI-AML-20 relies on the Read More …

Cloning an NSD3-Short-3xFLAG Construct into a Lentiviral Expression System

Antibodies are fundamental tools in the exploration of cellular biology. Unfortunately, there is a lack of high-quality NSD3 antibodies available. To overcome this, I have generated a bicistronic NSD3-Short-3xFLAG – IRES – PuroR lentiviral expression plasmid for the purpose of creating cell lines that stably express 3xFLAG-tagged NSD3-short. This will allow me to use a Read More …