A growing team of groundbreaking scientists around the world are now sharing their lab notebooks online
Hi, I would like to share with you the details for cellular assay detecting interaction between histone deacetylase HDAC6 and ubiquitin, which is a unique property within the histone deacetylase family. HDAC6 is a member of the histone deacetylase family which deacetylate lysine residues of histone and non-histone proteins. It plays an important role in Read More …
My previous results showed that overexpression of HDAC11 decrease basal acetylation levels of exogenous histones as well as decrease the acetylation increase caused by histone acetyltransferases. In order to determine if the observed effect is caused by catalytic activity of HDAC11, I used catalytic HDAC11 mutants Y304H and H142,143A (PIMID: 30819897, 28927261) and published HDAC11 Read More …
In my previous post I mentioned that there is controversy if HDAC11 can deacetylate histones. I decided to look at the effect of overexpression of HDAC11 on exogenous histone acetylation in cells. In my preliminary results I noticed that exogenous GFP-tagged histones, one day post transfection, have very low levels of acetylation; therefore I decided Read More …
Hi Everyone, This is the first time I am going to share with you my research and I will start with the project I am currently working on, which is the development of cellular HDAC11 target engagement assay. HDAC11, the only member of class IV HDAC subfamily, was believed to be responsible for the deacetylation Read More …
As a last step, to check if the assay qualifies for the screening purposes, Z’-factor was calculated. The dataset can be viewed on Zenodo.
For the purpose of compound library screenings, the compounds would be made available in DMSO and thus, when they would be incubated with the protein, the protein solution will now contain DMSO as well. It is important to determine the concentration of DMSO that could be tolerated by the protein such that the protein stays Read More …
The presence of detergent can prevent the sticking of the fluorophore to the sides of the wells of the plate and thus improve the signals. Hence, the effect of detergent was observed on the overall signals and the activity of HDAC11. The dataset can be viewed on Zenodo.
The linearity for HDAC11 activity for Boc-Lys-(TFA)-AMC substrate was monitored using the newly optimized buffer conditions. This was followed upon by Km calculation. The dataset can be viewed on zenodo.
An effect of adding reducing agent on the HDAC11 activity was monitored. Dataset can be found on Zenodo.
The pH range used earlier was expanded for further optimization to check if the activity could improve. The dataset is posted on Zenodo.
