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CODEX antibody conjugation

NL Ning Li
QZ Qin Zhu
YT Yuhua Tian
KA Kyung Jin Ahn
XW Xin Wang
ZC Zvi Cramer
JJ Justine Jou
IF Ian W. Folkert
PY Pengfei Yu
SA Stephanie Adams-Tzivelekidis
PS Priyanka Sehgal
NM Najia N. Mahmoud
CA Cary B. Aarons
RR Robert E. Roses
AT Andrei Thomas-Tikhonenko
EF Emma E. Furth
BS Ben Z. Stanger
AR Anil Rustgi
MH Malay Haldar
BK Bryson W. Katona
KT Kai Tan
CL Christopher J. Lengner
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Akoya antibodies were purchased preconjugated to their respective CODEX barcode (Supplementary Data 5). All other antibodies were custom conjugated to their respective CODEX barcode (Supplementary Data 5) according to Akoya’s CODEX user manual using the CODEX Conjugation Kit (Akoya, 7000009). Briefly, 50 μg of carrier-free antibodies (Supplementary Data 5) were concentrated by centrifugation in 50 kDa MWCO filters (EMD Millipore, UFC505096) and incubated in the antibody disulfide reduction master mix for 30 min. After buffer exchange of the antibodies to conjugation solution by centrifugation, addition of conjugation solution and centrifugation, respective CODEX barcodes resuspended in conjugation solution were added to the concentrated antibody and incubated for 2 h at room temperature. Conjugated antibodies were purified by 3 buffer exchanges with purification solution. 100 μl of antibody storage buffer was added to the concentrated purified antibodies.

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