ADD method for luminex

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Nathan Dwarshuis 2021-07-27 16:13:09 -04:00
parent dbf2869ae9
commit b541c280eb
1 changed files with 15 additions and 2 deletions

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@ -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 were removed from the retronectin plates using vigorous pipetting and
transferred to another 96 well plate wherein expansion continued. 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 \begin{table}[!h] \centering
\caption{Luminex Panel} \caption{Luminex Panel}