A growing team of groundbreaking scientists around the world are now sharing their lab notebooks online
Before I get into the details of my project and share my results, I would first like to give a brief background on HTT. What is HTT? HTT is a protein that is encoded by HTT, the gene, and it stands for huntingtin. This protein is heavily implicated in Huntington’s disease (HD), a disease Read More …
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 …
Our experiments use cells from patients with leukemia. These primary cells are notoriously difficult to culture, especially long term. To overcome this, we are using a co-culture system where we grow the leukemia cells with mouse stromal cells (OP9) to try to mimic some aspects of the bone marrow environment present in leukemia. Although essential Read More …
HDAC 11 protein was purified from Sf9 cells using Ni-NTA chromatography. The protein was concentrated to 1.4 mg/ml (above which it tends to precipitate) and flash frozen. The details on the experimental procedures can be found on Zenodo.
Histone decaetylases (HDACs) are one of the several enzymes that take part in regulating gene expression. They catalyze the removal of acetyl groups from ε-N-acetylated lysines from both, histone and non-histone protein substrates. By doing this, they maintain a balance with respect to the activity of the acetyltransferases in the cells. The HDAC family consists Read More …
Acute myeloid leukemia or AML is a type of cancer that is the result of abnormal division and development of blood cells. AML has very poor outcomes, with 5-year survival of less than 50 percent and even less in patients over the age of 60 [1]. Within AML, there are many subgroups that can dictate Read More …
Hi everyone! As part of SGC’s Extreme Open Science program, I’m going to be giving live updates on my project on the role of HTT in neurological disorders. As this is my first blogpost, I’ll start by giving an overview of my project. The focus of my project is HTT and its role in neurological Read More …
Every cell in your body contains the exact same set of genetic instructions encoded in its genome. However, for complex multicellular organisms such as ourselves, we depend on many specialized cell types that exhibit strikingly different features. This is largely accomplished by precisely controlling how and when genetic information is read out in each cell, Read More …
For the past few days, I have been working on optimizing a fluorescence polarization (FP) displacement assay for USP5 Zf-UBD. This assay will be used to test if small molecules can displace a fluorescence-labelled ubiquitin peptide from USP5. I started with screening different assay conditions with a small set of buffers. The method and data Read More …
Before I can begin characterizing the USP5 protein, I first needed to make some protein sample to work with. I expressed and purified the zinc-finger domain protein in bacteria and ended up with a high yield of a sample which is very pure. All of my methods and data from this experiment can be found Read More …
