Question: Is the Fixed RNA Profiling (i.e., Single Cell Gene Expression Flex) assay compatible with intracellular protein labeling?
Answer: Yes, the Fixed RNA Profiling assay is compatible with cell surface protein and intracellular protein labeling. If interested in labeling cells for cell surface protein or intracellular protein information with the Fixed RNA Profiling assay, users should refer to the Cell Surface & Intracellular Protein Labeling for Chromium Fixed RNA Profiling Demonstrated Protocol (CG000529). This document outlines cell surface and intracellular protein labeling protocols for use with Chromium Fixed RNA Profiling. General guidance on antibody-oligonucleotide conjugation, evaluating antibodies with flow cytometry, and best practices when staining cells is also provided here.
Both the cell surface and intracellular protein labeling protocols described in this document require a Fc receptor blocking step, after which the appropriate labeling protocol should be followed depending on the type of antibodies used. The Intracellular Protein Labeling Protocol, which includes a brief fixation before staining to allow cell permeabilization, can also be used with certain fixation compatible cell surface protein antibodies.
For details on sequencing Antibody libraries generated from PTG's MultiPro™️ Fixed Cell Immune Profiling Antibody Cocktail, see this article: What sequencing configuration is required for Proteintech Group (PTG)-derived antibody libraries generated from multiplexing experiments using Single Cell Gene Expression Flex?
For details on sequencing Antibody libraries generated from PTG's MultiPro™️ Fixed Cell Immune Profiling Antibody Cocktail mixed with Biolegend antibodies, see this article: What sequencing configuration is required for antibody libraries derived from using Proteintech Group (PTG) antibodies mixed with Biolegend antibodies, generated from multiplexing experiments using Single Cell Gene Expression Flex?
Products: Fixed RNA Profiling Gene Expression