Save hours on your immunoprecipitation experiments
Save hours on your immunoprecipitation (IP) experiments
Hi, everyone. Thank you for joining Proteintech and Chromotek's Tiny Technical Talk, on how to save hours on your immunoprecipitation. I'm Dr. William Olds, Scientific Officer at Proteintech Group.
So, as you know, there's so much to do. There's grants to do, there's manuscripts to figure out, and you just don't have time for your experiments.
And IPs particularly can take a long time, between from Cell lysate, all the way through Targeted antibody incubation, which lasts 1 to 12 hours, to Protein A/G bead incubation, which can also take 1 to 12 hours, all the way through washing and gel analysis of protein. IPs can take more than 3 hours and up to a few days.
Let's shorten that to 2 hours. Using Nano-Traps, which have been developed by Chromotek and Proteintech, this process can take less than 2 hours. Cell lysis to Nano-Trap incubation, which is using a Nanobody already pre-conjugated to a bead, this incubation can be less than 1 hour. And then from there, all you have to do is, wash it and allude it, and you can analyze your protein. All in all, this can take less than 2 hours, so less than your typical movie at this point.
What are Nanobodies? Nanobodies are the binding domains of heavy chain antibodies. Traditional antibody, thing here as depicted in the mouse, consists of two chains, light chain, and a heavy chain. However, in Alpaca, there are single-chain antibodies, and you can clone out the VHH, which is the binding domain of Nanobodies that actually bind to the protein of interest and you can conjugate those to beads to make what's called a Nano-trap.
Nanobodies are quite high affinity. As you can see in this gel here, during this incubation with this antibody, more of this band goes away, and it becomes very concentrated in what you've actually alluded from the Nanobody. And, we've determined that Kd of this antibody is 1 picomolar. That is just about the limit of how strong an antibody can be. And what this means is that, these results can be ready in less than 1 hour.
One of the ways that the Nano-trap can save you time, is that they are pre-conjugated and helping you in analysis, they have no heavy chains and light chains to take out and your western blot. These are already coupled to the beads, so you don't have to worry about doing an incubation with the beads like you would for conventional antibody. And this affinity is very strong, and so the protein remains on the beads, and the protein itself can remain on the Nanobody attached to the beads for a very long time, and even with very harsh conditions. Lastly, as I mentioned earlier, there is a single band that is in your analysis because you don't have to worry about the heavy chain and light chain like you do here at 25 and 50, all you see is your target protein.
We have quite a great Nano-trap portfolio, at the moment. We have a variety of Fluorescent tags here, some Solubilization tags like GST, some popular Peptide/Epitope tags, like Myc and V5, as well as a few Oncology Proteins.
To summarize here, GFP trap, you just have to lice, incubate and wash and allude, and the process takes less than 2 hours. However, for a conventional antibody, this can take more than 3 hours and even a couple of days. And what this means is, use these nano-traps, and you'll be just like this guy out here, enjoying the day with all the extra time that you have.
Lastly, I'd like to just talk a little bit more about Chromotek. Chromotek specializes in Nanobody-based Reagents for elevated performance. They specialize in immunoprecipitation and affinity purification of proteins, antibody capture, immunofluorescence, super-resolution microscopy reagents, and reagents for life cell imaging. Website as shown here. It was founded in 2008 as a spinoff from a Ludwig Maximilian at the University of Munich, Germany, using a proprietary technology, in order to develop the Nanobodies. And, they've had over 20,000 customers, in 50 countries, and there are more than 2,600 citations of the nano-traps within the academic literature. And so with that, I want to thank you, and you can order online at pglab.com or chromotek.com, and I hope that you have more time to do what you want with your time.