Question: Is the 3’ gene expression profiling assay compatible with TotalSeq™-B Intracellular Protein Staining?
Answer: Although intracellular staining with TotalSeq™-B antibodies in the Single Cell 3’ gene expression Solution has not been tested or validated by 10x Genomics and is not supported, limited guidance can be provided to help enable these experiments following BioLegend protocol.
This 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. Please contact BioLegend for additional assistance with the assay.
Furthermore, 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. We also provided limited additional guidance as a starting point to help enable intracellular staining in the Fixed RNA Profiling Solution. More information can be found in this knowledge based article: Is the Fixed RNA Profiling assay compatible with intracellular staining?
Please note, intracellular staining is not officially supported by 10x Genomics.
Products: Single Cell Gene Expression, Fixed RNA Profiling, Single Cell Gene Expression Flex