This protocol is used to determine the optimal oligonucleotide-conjugated antibody concentration to spike into the 10x Antibody Panel.

This protocol optimizes the concentration of a single, oligo-conjugated antibody. Antibodies can be purchased from BioLegend or generated through custom conjugation approaches. 10x Genomics does not support the process of independent conjugation of antibodies for Visium applications.

Repeat this protocol when optimizing multiple antibodies. This protocol also provides guidance on pooling optimized antibodies and includes a Pooling Workbook (CG000693) for additional assistance. For a discussion on using immunofluorescence vs. sequencing to analyze protein data, refer to Antibody Optimization - Key Considerations within this Demonstrated Protocol.