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Simple Western and Single-Cell Western have the potential to make significant contributions to three important branches of immuno-oncology: angiogenesis of the tumor microenvironment, immune checkpoint inhibition, and chimeric antigen T-cell (CAR T-cell) therapy. Learn more.
In this Application Note, we show how to achieve ultrasensitive total protein detection using the 5X biotin labeling reagent with a focus on AAV analysis.
Here, we present data showing the power of using automated Simple Western platforms to screen panels of Degrader and IMiD compounds in order to quantify degradation activity. In this study, we demonstrate the time savings achieved by automating these large screens as well as Simple Western’s ability to accurately quantify DC50 and Dmax values for specific Degraders and IMiDs.
A multi-antigen serology assay for COVID-19 using Simple Western furthers understanding of the COVID-19 disease progression and the efficacy of a vaccine.
In this Application Note, we demonstrate how Simple Western can be applied for the detection and characterization of ACE2 and TMPRSS2, key players in SARS-CoV-2 infection. As an open platform, any commercial or custom antibody may be used for target protein detection on Simple Western. Here, we used an anti-ACE2 antibody from R&D Systems and an anti-TMPRSS2 antibody from Novus Biologicals to develop immunodetection assays for these proteins in purified form and expressed in human cells. This resulted in assays that were highly sensitive over a large dynamic range, providing molecular weight information and even quantification in a human cells.
To gain the most data out of your precious sample, Jess provides a two-step immunoassay that is
performed within the same capillary. This new feature, called RePlex removes the antibodies from the
first round of probing to perform either a second round of probing with new antibodies or total protein
detection. Importantly, RePlex efficiently removes antibodies between probing without compromising
the integrity of the immobilized protein or its epitopes, allowing for excellent reproducibility across
cycles. With Jess’s chemiluminescence and fluorescence channels, you can detect multiple targets per
cycle. The second cycle can also be dedicated to total protein detection so that you can normalize your
data with confidence. All the steps of RePlex are automatically performed with Jess, providing more data,
and lowering the cost of reagents and consumables per sample.
In this application note, we’ll show you how Wes™ and Milo™ partner to get you critical answers to 1) what type of immune cell populations are present in a sample and then 2) what percentage of cells in that sample make up a specific immune cell subtype.
In this application note, you’ll see how CGTC has used highly-specific antibodies exclusively manufactured by PROGEN with fully automated Simple Western™ assays on Wes™ to monitor and characterize AAV capsids during product purification.
In this application note, we simultaneously assess p-Akt and total Akt levels using different detection channels combined with in-capillary protein normalization to demonstrate the superplexing power Jess possesses.
シンプルウェスタンTotal Protein Assayの同時測定で、イムノアッセイデータのノーマライゼーション
In this application note, we focus on the accurate detection of four candidate contaminants that may be present during various stages of the therapeutic protein and vaccine development processes: host cell protein (HCP), Protein A, green fluorescence protein (GFP) and bovine serum albumin (BSA).
In drug discovery, confirmation of in-cell target engagement is a critical component of the drug development process. Confirming that a drug candidate engages its proposed target in the cell and determining the concentration at which it exerts the desired effect(s) fulfill fundamental criteria for translation to activity and efficacy in its target tissue. Thermal shift assays (TSA) are regularly used by industry and academia to uncover or confirm interactions using purified proteins. Recently, this type of assay has been adapted to a cellular format and is called the cellular thermal shift assay (CETSA). In this application note, quantitative and reproducible CETSA data generated with ProteinSimple’s Wes instrument (CETSA-Wes) are presented. This assay verifies drug target binding, and given its quantitative nature, the half maximal inhibitory concentration (IC50) is also calculated.
In this how-to-guide, we'll show you how to use the Total Protein Detection Module (DM-TP01) with any immunoassay detection module of your choosing to get total protein and immunoassay data in the same run.
In this application note, we’re honing in on the biomarker verification and validation steps. Proof-of-concept data was generated to demonstrate how Simple Western and Simple Plex assays data give similar trends and work together to give you fast, sensitive, and precise information about your biomarkers of interest.
Scientists measure serum antibody levels to confirm immune responses against a bacteria to diagnose infections, to test for antibody production after vaccination, and to detect the presence of autoantibodies in autoimmune diseases. Traditional Western blots are often used to detect these antibodies, but testing with Western blots means a lot of hands-on time. After the antigen of interest is separated by SDS-PAGE and transferred to a membrane, each lane has to be cut into individual strips so patient serum samples can be individually tested for the presence of specific antibodies. Then you have to process and analyze them manually.
Simple Western assays happen in individual capillaries, and everything from sample separation to data analysis is completely automated. No more cutting individual strips, washing and incubating them, or lining them all up with a molecular weight marker before detection. Just pipette your sample into the wells of your assay plate, set up your run, and you're done! And all that manual data analysis is gone too — Compass for Simple Western does it all for you. Did we mention you only need 10 µL of diluted serum per data point? That means you'll get a lot more data points for every 1 µL of neat serum.
In this application note, we used Simple Western to detect autoantibodies in lupus patient serum as a model system to generate proof-of-concept data for the assay. But you can use this method any time you need to detect and quantitate specific serum antibodies. In fact, check out how researchers are using this assay to detect Salmonella antibodies without having to cut blots into individual strips.
Laser capture microdissection (LCM) is a powerful tool to identify and isolate a pure sample of the cell type you're
interested in. But, proteomic studies with LCM samples are really restricted by the small amounts of tissue collected with
each capture since there just isn't much to work with. You often have to use the entire sample captured for traditional
Western blot analysis and that only leaves you with one data point! So, researchers often use 2D Electrophoresis and
mass spectrometry instead to max out the amount of data they can generate. Both of these methods have their own
limitations when it comes to ease-of-use and reproducibility though. Immunohistochemistry is also used to analyze LCM
samples as it's a more accessible technique, but it really doesn't give you a lot of info either.
Simple Western is an automated capillary-based immunoassay that changes the proteomic research game. You only
need 1-10 µL of LCM sample for each data point, so you'll get more data points for each sample you collect. Not to
mention the sensitivity that comes with it will even let you analyze proteins you couldn't previously do with traditional
Western blot. And it's all wrapped up in a simple workflow that minimizes your hands-on time. Simple Western is a
sensitive, easy-to-use analytical tool that ups the ante on protein analysis with LCM samples.
In this application note, we'll show you two examples of how Simple Western changed what researchers could do with
their precious LCM samples for the better
The Simple Western Immunoassay is the gel-free, blot-free and hands-free solution for researchers looking for a better way to get their Western blot data. The simple fact that you get analyzed data in just three hours with only 30 minutes of hands-on time changes things forever!
Peggy enables researchers to follow up a size-based immunoassay with a charge-based assay on one platform using the same sample. The charge-based analysis provides an information-rich, complementary data set, elucidating ratios of protein variants and providing leads for biomarker development. Like other Simple Western products, Peggy provides a fully automated solution, from loading samples
all the way to peak analysis.
Wes takes the benefits of Simple Western assays a step further by simplifying their workflow with pre-filled plates and disposable capillary cartridges. This application note gives an overview of how to transfer a traditional Western blot assay to Wes.
RNA interference (RNAi) is an RNA-dependent gene silencing mechanism that can affect the expression of specific genes by inhibiting translation or suppressing transcription epigenetically. Using Firefly assays, RNAi effects such as impact on phosphorylation or silencing can be studied functionally in samples as small as 100 cells. An additional benefit of the small samples size is that a variety of conditions can be studies in a single assay.
Monophospho- and diphospho-ERK isoforms are not resolved by traditional Western blot analysis, and the sample quantity required is relatively large. The Firefly ERK1/ERK2 assay can distinguish and quantify unphosphorylated, mono and dual-phosphorylated isoforms of ERK1 and ERK2, allowing a more accurate determination of ERK activation than Western blots provide.