Advanced Western Blotting Solutions for Neuroscience

Table of Contents

• Get the Most Data from Your Precious Samples
• RePlex with Total Protein Detection
• Industry-Leading Fluorescent Detection Sensitivity with Stellar 
• Dissociation of Mouse Neural Tissue for Single-Cell Western Analysis
• Identify and Quantify Single Neural Cell Subtypes
• Fully Automated Westerns Enable Cutting-Edge Neuroscience Research
• Advances in Neuroscience
• Hear from Your Peers

The Most Data from Precious Samples

Some of the most prevalent and least understood diseases are those that affect the nervous system. Simple Western™ is winning the race to discovery in neurological diseases by offering high sensitivity quantification of proteins from rare, sparingly-available samples at unprecedented speeds while also offering multiple multiplexing strategies to get the most data out of your precious neuroscience sample. With Single-Cell Western on Milo™, you can measure heterogeneity in your samples and study protein expression within specific rare neural cell subtypes.

Simple Western requires only 3 µL of sample while offering pg-level sensitivity, so there is no longer any need to pool samples like mouse tissue. Get single-animal statistical data and realize significant cost savings and save mouse lives. Simple Western also ups the data point count on laser capture microdissection (LCM) samples.

RePlex with Total Protein Detection

To get the most data out of precious samples, Simple Western also eliminates the arduous strip and re-probe procedure by offering RePlex™, which efficiently removes the antibodies from the first round of probing for a second round of probing with fresh antibodies. The second round may also be used for Total Protein Detection, so you can normalize your data with confidence.

Right Figure: Total protein concentration was used as the internal loading control for protein quantification in tau transgenic mice. (A-B) Simple Western analysis of brain homogenates from control or tau transgenic mice using antibodies against Lamin A/C and b-Actin. (C) Simple Western Total Protein assay to detect total protein in each sample. (D) Quantification of Lamin A/C, (E) b-Actin and (F) total protein concentration. (G) Representative electropherogram generated from the Simple Western Total Protein assay. Figure adapted with permission from Musi et al. (2018) Aging Cell. 

Industry-Leading Fluorescent Detection Sensitivity

As the current flagship of the Simple Western product line, Jess™ provides users with the most flexibility in protein detection with sensitive chemiluminescence and NIR/IR fluorescence channels. Now NIR/IR fluorescence detection on Jess is even better with Stellar™ NIR/IR Detection Modules, which provides industry-leading fluorescence sensitivity at detection levels below 1 pg, along with excellent reproducibility and a 4-log dynamic range. With this leap in sensitivity, Stellar fluorescence detection rivals the widely recognized sensitivity of chemiluminescence detection and demolishes the fluorescence detection of the closest competing traditional Western blot imaging technology, which requires at least 50 pg.

Dissociation of Mouse Neural Tissue for Single-Cell Western Analysis

In this App Note, we describe and characterize a protocol to successfully dissociate mouse neural tissue micro-surgically dissected from combined cortex, ventricular zone, and hippocampus regions of E18 mice into single cells. Using a multiplexed Single-Cell Western assay to analyze protein expression in the dissociated single neuronal cells, we identify 3 subpopulations of cells within the tissue: astrocytes, radial glia, and immature neurons based on expression of their respective phenotypic protein markers: GFAP, Pax6, and β-III tubulin.