RNA Immunoprecipitation (RIP) is growing up. After suffering through some awkward adolescent years, RIP has evolved from its humble beginnings in a few hardcore labs and is now stepping out shadow of its more famous cousin, Chromatin Immunoprecipitation (ChIP).
The more mature RIP has been steadily building its fan base too with curious researchers seeking to snag glimpses into the RNA world by studying coding and non-coding RNA mechanisms, and other ways non-coding RNAs interact with cellular machinery. Now with recent refinements, it’s easier than ever to run a great RIP experiment. We chatted up the Millipore R&D team, who happen to appreciate RNA as much as the other nucleic acid, to get the low down on how those latest advancements have helped push RIP technology towards adulthood.
RIP’s Troubled Past
When we first wrote about “The Rise of RIP” in 2008, the method was still going through some growing pains. Even though it was the RNA version of ChIP, with similar features; like using antibodies to isolate binding proteins (RBPs instead of chromatin complexes), RIP never got the same attention as ChIP. But as researchers broadened their affection beyond the usual suspects (e.g. transcriptional regulation), protein-RNA interaction interest increased and RIP got some legs under it.
The absence of specific materials and validated reagents left epigenetic researchers to work out RIP’s kinks for themselves though, forcing them into making and optimizing their own reagents. Qualified RNA antibodies? A few years ago these were rarer than a Nellie sighting. This of course made RIP experiments difficult and temperamental to run.
RIP put it’s checkered past behind it. Hearing the pleas of RIP enthusiasts, companies are offering products tailored just for RIP that eliminate most of the dirty work and let anyone quickly become the local RIP expert. “With our RIP Product line, we are really looking to enable this emerging area of epigenetics.” Says Michael Sturges, PhD, Sr. Product Manager for Epigenetics at EMD Millipore. “RIP can be used to explore post-transcriptional gene regulation, as well as transcriptional gene regulation, depending upon the RNA binding protein target of interest”.
RIP Antibodies Get Filled Out
Like getting 6-pack abs in the gym (something we know little about, but are happy to use the analogy), making/validating a great antibody doesn’t happen overnight. So it’s no surprise that RBP specific antibodies were in short supply for a long time. Lately the list of RBP antibodies is growing by leaps and bounds, and some, like EMD Millipore’s RIP Ab+ antibodies, even come RIP validated. “Every lot of our RIP Ab+ antibodies gets stringently QC’d… in the RIP application. Plus, a negative control antibody and control primers are included to ensure the best possible performance for researchers.” Explains Sturges.
That sounds like a much easier way to get good abs than endless crunches to us.
RIP Leaves Home(Brew)
It happens. Methods grow up, mature and eventually leave home…the homebrew that is. Like many other molecular biology methods that originated in the labs of talented researchers, RIP has now migrated into a more commercial setting. Don’t call it a sell out though. The new kits on the market are less juvenile, and set up so that even your undergrad intern won’t mess them up.
For example, Millipore’s EZ-Magna RIP Kit comes complete with reagents set up to make RIP virtually plug-and-play. “We use optimized Protein A/G beads for a wider selection of compatible antibodies, RNAse inhibitors and RNAse-free components, and include negative and positive controls to make RIP something that any lab can easily adopt.” Sturges adds.
With all of the dedicated antibodies and reagents out now, it looks like RIP is finally primed for maturity.
Check out the fully grown RIP Products at the EMD Millipore website.