Scientists Test Histone-modification Antibody Quality.

Scientists all over the world have been busy figuring out how DNA is packaged in cells, because a stretched out DNA strand would be as long as sic feet. But it fits nicely packaged inside a nucleus of one of our cells.   The packaging comprise an amino acid molecule called a histone, mostly and influences the on and off switches of different genes that regulate cellular function and play a role in human diseases ranging from cancer to genetic disorders.

Scientists study histones by using antibodies to specific “flavors” of histones that are only very slightly different from one another. The antibodies help to pinpoint what DNA is being packaged by a certain kind of “flavor” of histone, and how that affects gene regulation. Different flavors affect genes differently.

In a paper published today in the journal Nature Structural and Molecular Biology, Jason Lieb, PhD and his colleagues from across the country describe how they tested more than 200 antibodies against 57 histone  flavors in three different organisms, using three different tests commonly used in this kind of genetic analysis.  They found that about 25 percent of antibodies currently sold have a problem with specificity – targeting the anticipated histone – in a given test.  They believe that this proportion is likely to remain steady over time.

“And this is where it gets complicated, many companies make these antibodies that we scientists use in our labs – but there are so many different kinds of histones and types of tests we do that it’s just not feasible for the companies to anticipate every single way that a given antibody can be used. Histones are essentially the key to the DNA library.  They tell you which ‘shelves’ of that library – or areas of the genome – are open or closed to information moving in and out.  But since the differences between the different ‘flavors’ of histones are often extremely small, and it’s likely that an antibody may react with more than one histone or in different ways depending on the type of test being used in the lab.  It makes scientific precision very difficult. So we thought, OK, we need to help ourselves as scientists.  We set up a web-based searchable database at http://compbio.med.harvard.edu/antibodies.  Our results are there and other scientists can also post their results so that we have a self-sustaining, up-to-date source of information that is really important to scientists working to understand a broad range of genetic phenomena,” says Jason Lieb, PhD, ho is a professor of biology at the University of North Carolina at Chapel Hill and member of UNC Lineberger Comprehensive Cancer Center, and who led the project testing common antibodies.

The research was funded by the National Human Genome Research Institute (part of the United States National Institutes of Health) and included researchers from the Universities of California at Santa Cruz, Berkeley, and San Diego, the Lawrence Berkeley National Laboratory, the Ludwig Institute for Cancer Research, Harvard Medical School, the University of Cambridge (UK), Washington University in St. Louis, Ontario Institute for Cancer Research (Canada) and Rutgers University.
UNC Press Release