Simple Western technical library

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.

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.

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, 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.

Immunotherapy has opened the door to a new era in cancer treatment where dramatically successful and durable responses are being reported. Still, the population of patients who respond are a minority, and this highlights the need for a deeper understanding of the immune response to cancer. To that end, accumulating evidence suggests that the heterogeneous makeup of the tumor microenvironment (TME) is a major contributor to the discrepancies in response observed.

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

Simple Westernでバイオプロセス中の不純物質の検出
医薬品製造過程関連の不純物の同定および定量

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 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.

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.

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.