From b541c280eb94b2e496b41695dc4ce0545472d2f9 Mon Sep 17 00:00:00 2001 From: ndwarshuis Date: Tue, 27 Jul 2021 16:13:09 -0400 Subject: [PATCH] ADD method for luminex --- tex/thesis.tex | 17 +++++++++++++++-- 1 file changed, 15 insertions(+), 2 deletions(-) diff --git a/tex/thesis.tex b/tex/thesis.tex index 102bf0d..83b4744 100644 --- a/tex/thesis.tex +++ b/tex/thesis.tex @@ -931,9 +931,22 @@ plates and incubated for another \SI{24}{\hour}. Cells and beads/\glspl{dms} were removed from the retronectin plates using vigorous pipetting and transferred to another 96 well plate wherein expansion continued. -\subsection{Luminex} +\subsection{Luminex Analysis} -% METHOD luminex +Luminex was performed using a \product{ProcartaPlex kit}{\thermo}{custom} for +the markers outlined in \cref{tab:luminex_panel} with modifications (note that +some markers were run in separate panels to allow for proper dilutions). +Briefly, media supernatents from cells were sampled as desired and immediately +placed in a \SI{-80}{\degreeCelsius} freezer until use. Before use, samples were +thawed at \gls{rt} and vortexed to ensure homogeneity. To run the plate, +\SI{25}{\ul} of magnetic beads were added to the plate and washed 3x using +\SI{300}{\ul} of wash buffer. \SI{25}{\ul} of samples or standard were added to +the plate and incubated for \SI{120}{\minute} at \SI{850}{\rpm} at \gls{rt} +before washing analogously 3X with wash. \SI{12.5}{\ul} detection \glspl{mab} +and \SI{25}{\ul} \gls{stppe} were sequentially added, incubated for +\SI{30}{\minute} and vortexed, and washed analogously to the sample step. +Finally, samples were resuspended in \SI{120}{\ul} reading buffer and analyzed +via a Biorad Bioplex 200 plate reader. \begin{table}[!h] \centering \caption{Luminex Panel}