Western analysis directly in your cells? YES you can!
Ever heard about doing a Western in your cells? It’s new, it’s easy, it’s fast and ..... it’s here!
Finally you have the possibility to perform westerns in your cells: there is no need for protein extraction and endless blotting procedures. You can quantitate protein levels directly in your cells with In-Cell Western assays. What’s more, you can easily analyze at least 96 samples in one go.
Detect proteins in fixed cultured cells
I bet you’re curious how In-Cell Western assays work. Similar to when you prepare your samples for conventional western blotting, you seed your cells in culture plates and, if desired, treat them appropriately. After incubation, you wash, fix and permeabilize your cells. Instead of performing protein extraction from here, you immediately incubate your cells with primary antibodies and subsequently with IRDye secondary antibodies. These secondary antibodies are used for near-infrared (NIR) detection with the Odyssey systems from LI-COR. NIR technology eliminates background fluorescence from other cell components so you can easily and accurately quantify protein levels. Place your plates in an Odyssey system and let the quantification begin while you have yourself a coffee…
Quantify two targets at once
Instead of splitting your protein extracts in multiple samples or re-probe your western blots to analyze more than one target, with In-Cell Westerns you can detect total protein and phosphorylated protein, or two any other targets, in the same sample in one go; one at 700nm and one at 800nm. Did I tell you already that In-Cell Westerns are so easy?
Easily quantify 96+ samples in one go
I have mentioned it earlier: analyzing over 96 samples in one go has never been easier. Since protein analysis is done directly in culture plates, you can quantify protein levels in a lot of samples. Take it up a notch and use a 384-well plate; it won’t increase your analysis time dramatically. Make it even more enjoyable by using a multichannel pipet to make sure you experience the easiest and fastest Western experiments.
So for the lucky Odyssey owners out there: there is no need to keep on performing those endless Western blots to quantify protein levels. Once you’ve tried this easy and fast Western analysis, you never go back.
For those of you that don’t own an Odyssey system yet: learn more about the sensitivity of NIR imaging and experience easy Western analysis the fast way.