My School and Department are in the process of purchasing a Luminex 200 system for detecting and quantifying targets from plasma/serum and cell culture.
I would very much appreciate any opinions, suggestions and insight others may have with regards to using this technology for targets extracted from brain tissue.
I realize the commercially available multiplex assays offered are limited with regard to specific CNS targets.
Thanks in advance, Scott
Yes, you are going to have to do some work in trying to use commerically available kits that usually are not tested for any other samples but plasma/serum or supernatants.
Depending on how precious the samples are, and your budget, you can customize a commerical kit to your samples. Below is a published article, using (not the Luminex 200) a bead based assay that you can use as a starting point.
Article Optimization of multiplexed bead-based cytokine immunoassays...
For the most academic research labs, most people just use the protocol for serum/plasma. It will work. More extensive fine tuning to control for matrix effects is probably beyond the scope of this topic.
We frequently use luminex based assays in our lab to assess cytokine expression. The assay is very consistent and I would definitely recommend this initiative by your lab towards investing in the equipment.
Hi,
I have experience in Luminex for cytokines and quimiokines in human Broncho-Alveolar Lavage concentrated, culture supernants as serum. It works very well. Be
Hi Scott,
I routinely run brain tissue lysates on a Luminex Flexmap3D (high-through put version the Luminex 100/200). I use a lysis buffer from ThermoFisher/Procartaplex (you can prepare it yourself, recipe is provided in their technical data sheet). After homogenizing the brain tissue, you will need to centrifuge at 18,000 g for 20 mins at 4degrees C. Note: all steps require processing at 4C and avoid freeze-thawing! You will need to perform a Bradford assay and adjust the total protein to 10 mg/ml (some Luminex kits require a different protein concentration). If you can’t run the assay the same day as processing the tissues, aliquot into low protein binding polypropylene tubes and freeze at -80C. Upon thawing your samples (on ice), spin the lysates at 18,000g for 10mins at 4C, then pipet into your prepared beads and follow the kit instructions.
Diane
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