Having alternatives is always great, particularly when it comes to studying histone modifications. By using some recombinant proteins of histone reading domains, researchers from the lab of Albert Jeltsch at Stuttgart University (Germany) recently demonstrated that these domains might just one-up antibodies when it comes to deciphering the combos of the histone code.
Lead author Goran Kungulovski shares that “Histone modification antibodies have been the workhorse of chromatin biology, but recently numerous studies have raised concerns about their insufficient specificity and high batch to batch variability. In our study we introduced and experimentally validated the idea that Histone Modification Interacting Domains – HMIDs (also popularly termed ‘reading domains’) can be used in lieu of histone modification antibodies in many chromatin biology approaches such as chromatin precipitation coupled with qPCR or deep sequencing and western blot.”
The technology allows for a cost-effective production of recombinant reading domains with high yields in E. coli, but that’s also true for recombinant antibodies.
However, the advantages of using ‘reading domains’ over antibodies are aplenty, including:
- Consistent quality and reproducibility due to rock solid invariable properties across different lots.
- The domains only need to pass quality control once (compared to batches of antibodies which have to be validated and documented individually).
- Their modular design makes them more fun than lego, and with similar properties too, since clever engineering can be used to create a number of other great features, including:
- Domains with novel single or dual specificities.
- The addition of affinity tags and matching negative (pocket mutant) controls, which are a level of control that can’t be achieved with antibodies.