Stunner

Hi. Hey. Howdy. Oh, are we rolling? Hi. I'm Brian, and I'm here to introduce you to measuring collateral stability with Stunner, the ultimate quantification and sizing tool. Oh, yeah. Stunner is the only plate based platform that bundles UV Vis and light scattering to get you k d, b two two, concentration, and tons more deeds on your proteins from just two microliters of sample. When screening your formulations, you wanna make sure your proteins are in the right environment and check if they're at risk for aggregation by seeing how they interact with each other. Measuring k d b two two for a protein in different formulations lets you know if your proteins will continue to be happy hanging out side by side even when concentrations go up and things get crowded. The current methods use too much volume, can't keep up with high throughput needs, or don't measure concentration at the same time. Let's see how Stunner here gets it done. We'll start off by loading our Stunner plate. Just pipette in two microliters for each sample in a dilution series and the microfluidic channels pull your samples in. Once your sample is ready to be read, it gets pulled through two microcubets with fixed path lengths to ensure the most accurate and precise UV Vis concentration data and light scattering data from the smallest possible volume. Now let's jump into the Stoner software. We'll select the protein button and select the application for KD and B22. Give your experiment a name and we're set. Here we can create our plate layout either manually by adding in our blanks and samples or we can import from an excel file. Select whatever type of protein you have, the buffer you're working in, and stunner is ready to go. And now we just drop in our plate and stunner is gonna turn up KD and B22 in no time. In stunner's analysis software, KD and B22 are served up for you right from the start. Positive slopes in the KD and b two two data are what you wanna see and mean that your protein is in the right formulation. If slopes are negative, watch out since that means attractive self interactions are putting your protein at risk for aggregation. If you want, it's easy to drill down into the data and also grab underlying concentration and size info. Now that you've seen how you can easily measure k d b two two and look good while doing it, you're set for hassle free colloidal stability of your proteins. Guys, is anyone still here?

Steady. Steady. Oh. Hi. I'm Brian, and I'm here to introduce you to protein quant, contaminant, and aggregation detection on Stunner, the protein quantification, sizing, and aggregate detection workhorse. Stunner is the only plate based platform that bundles UV/Vis and light scattering to give you accurate protein quant, aggregation detection, and tons more deets on your proteins from just two microliters of sample, and flexes Unmix algorithms that read out contaminants from your results to give you insights as to what's hiding in the mix and to dish up next level accuracy. When working with proteins, aggregates and contaminants can be lurking in your samples, completely throwing off accuracy of your measurements and risking you sending bad candidates to the next step. So getting accurate measurements is crucial to avoid getting burnt down the line. But traditional methods don't account for other components in your sample that could be contributing to this absorbance, such as turbidity or unwanted impurities. Stunner takes that UV/Vis absorbance spectrum and uses intelligent unmixed algorithms to separate out different components contributing to that total spectrum. Unmixed not only tells us if we have impurities in our samples, but also gives us an accurate protein concentration by only measuring the absorbance at two hundred eighty nanometers that's coming from our protein. Let's see how Stunner rolls it all out. Oh, so that's what a cupcake is supposed to taste like. Good job, buddy. We'll start off by loading our Stunner plate. Just pipette in two microliters of each sample and the microfluidic channels pull your samples in. Once your sample is ready to be read, it gets pulled through two microcuvettes with fixed path lengths to ensure the most accurate and precise UV/Vis concentration data and light scattering data from the smallest possible volume. Jumping into Stunner software, we'll select the protein button and select the application for proteins, lysates plus sizing to unleash all of Stunner's skills for measuring concentration, impurities, sizing, and aggregation on each sample. Impurities that are identified in these samples are listed in the app description. For protein samples, we've baked in some of the most common impurities for Stunner's Unmix to separate out. If you're working with samples from a different source, no worries. Stunner has apps designed for different sample types and their most common contaminants. Now we give our experiment a name, and we're set. Here, we can create our plate layout either manually by adding in our blanks and samples, or we can import from an Excel file. Select your protein type or added protein from the Analyte pull down list and the buffer you're working in, and Stunner is ready to go. And now we're just gonna drop in our plate, and Stunner's gonna cook up quant and aggregate detection in no time. Here we go. Looks like the results are ready. Now that we've whipped up some delicious results, we can review them starting here at the overview page. Here, we can see the summary of each sample with the absorbent spectrum displayed for each well. Moving over to well details, we can chew on the details from each well individually. All replicates are grouped here, so we can do a quick reproducibility check on our samples by selecting the sample instead of an individual replicate. When individual wells are selected, we can keep an eye on contaminants that are present in that sample and how much they are influencing the absorbance at two hundred eighty nanometers. To unwrap the unmixed spectrum, let's start by looking at the black line, which is the total UV/Vis spectrum. The green line is the portion of the spectrum from protein in our sample, and the blue line is the contribution from our impurities. To assess the fit, the yellow line represents the deviation from an exact fit. Here we can see the concentration from traditional UV/Vis and Unmix are very similar, which tells us that the samples are pure with minimal contaminants contributing to the absorbance at two hundred eighty nanometers. Whereas for this sample, Unmix flags significant contamination and gives us a breakdown of each of the components to give us our true concentration value. Finally, along with the unmatched results, sizing results from DLS are also collected on the same sample. We can see the average size of each sample and also whether aggregates are hiding under the main course. Now that you've seen how you can easily measure quant and aggregate detection on Stunner, you're ready to pick your best of the batch in no time. That is just delicious. Okay. How does this one taste? Maybe you should just stick to science. No.