Question: Is the 3’ gene expression profiling assay compatible with TotalSeq™-B Intracellular Protein Staining?
Answer: Intracellular staining with TotalSeq™-B antibodies for use with the Single Cell 3’ Gene Expression assay has not been tested or validated by 10x Genomics and is therefore not supported.
The protocol for detection of intracellular antigens requires workflow modifications and PFA fixation which is not supported for 3’ assays. PFA fixation is expected to negatively impact Gene Expression data quality. Additional optimization of staining and wash buffers may be required. For limited guidance on the workflow modification please contact Biolegend.
If interested in intracellular protein labeling, we recommend customers use our Single Cell Gene Expression Flex (Fixed RNA Profiling) assay, which was optimized for use with formaldehyde-fixed samples and shows robust data quality across a range of sample types. With the Flex assay, cells can be labelled for cell surface protein and/or intracellular protein information with the Fixed RNA Profiling assay. If interested in this application users should refer to the Cell Surface & Intracellular Protein Labeling for Chromium Fixed RNA Profiling Demonstrated Protocol (CG000529).
Products: Single Cell Gene Expression, Fixed RNA Profiling, Single Cell Gene Expression Flex