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[Music] look closer Milla Plex is the largest portfolio of verified assays for use on Luminex instruments each assay is configurable to run the analytes you want all assays offer reliable performance across Lots we offer more than eleven hundred analytes arranged in configurable panels by application you have the freedom to choose only the analytes you want all Milla Plex assays are powered by X map multi analyte profiling technology licensed from Luminex corporation each Milla Plex assay contains all reagents necessary to run 196 or 384 well plate the core X map technology uses color-coded microsphere beads beads compatible with the Flex map 3d contain a third dye resulting in up to 500 distinct bead region possibilities beads are selected and coated with specific capture antibodies ultimately forming a sandwich immunoassay specific for detecting an analyte of interest Milla flex assays are capable of detecting up to 48 different analytes in the same sample using unique bead sets for each analyte Milla Plex assays save you time compared to a traditional Eliza with 40 samples run in duplicate and 10 target analytes which may take 10 hours to perform by Eliza Milla Plex takes just four hours and uses a fraction of the sample while offering a comparable level of sensitivity a cost savings is realized by combining multiple analyte assays into a single kit look closer let's review the Milla Plex assay protocol and highlight some key steps the protocol for the analysis of soluble biomarkers is very similar to performing a sandwich Eliza and the entire assay can be completed within a single day or with an overnight incubation in between if desired for maximum sensitivity simply follow the plate map template located in the back of each protocol in general 76 wells are available for samples or 38 samples can be run in duplicate now let's highlight key steps in the protocol add standards and samples to plate wells and then add conjugated Milla flex beads to all wells as indicated in the protocol be sure to use well calibrated pipetters throughout the protocol Sonic eight each bead file individually and then vortex as instructed in the protocol all beads are combined in the provided mixing bottle we also offer premix bead options for selected panels beads are coated with antibodies or proteins to capture specific analytes in all Miller Plex assays when beads are mixed with sample the analytes are captured by the specific antibody or antigen coding the bead perform a two-hour incubation at room temp with shaking or incubate samples overnight at 4 degrees Celsius to maximize sensitivity carefully wash beads to remove any unbound sample add detection antibody cocktail and then ese PE wash again to remove any unbound fluorescent reporter a fluorescent reporter tag conveys the quantitative results for each analyte now let's see how our scientists do it in the lab perform 3 washes for 5 minutes each washes may be performed on an automated magnetic system such as the biotech plate washer shown here or manually with a handheld magnetic plate washer after washing incubate with detection antibody for one hour at room temperature next perform three more washes for five minutes each then incubate with PE labeled streptavidin for 30 minutes at room temp remember that beads and SI PE are light-sensitive finally perform a final set of three 5-minute washes run the plate on the luminex instrument with exponent software luma Nexus flex map 3d has a fast read time and compatibility with both 96 and 384 well plates the lumen X 200 and Flex map 3d have high flexing capabilities compatible with higher throughput applications mag pix is an affordable entry-level option for labs with budget constraints as a specifications are set during the assay design phase by our team of research and development scientists all Milla Plex assays are verified to fit established millets assay development criteria and provide consistency in our portfolio offering ISO 9001 quality systems mitigate deviation during reagent manufacturing curve potency is checked for each analyte in each new lot to a shifting in the standard curve and inconsistent sample readings between locks our assays are for research use only and not intended to be used for clinical or for medical diagnostic purposes our immunoassays portfolio includes human primate rat mouse canine and a variety of other species if you are not using Milla plaques assays or Luminex instrumentation would you like a live demo get started by contacting one of our field application scientists today by visiting sigma-aldrich comm slash Milla Plex [Music]