Cell Surface Protein Labeling for Single Cell RNA Sequencing Protocols

Demonstrated Protocol, Last Modified on March 31, 2021, Permalink

CG000149_DemonstratedProtocol_CellSurfaceProteinLabeling_RevC.pdf

CG000193_Barcode_Whitelist_forCustom_Feature_Barcoding conjugates_RevA.txt  (right-click and Save Link As...)
CG000193 Barcode Whitelist forCustom Feature Barcoding conjugates RevA

Cell surface proteins can be labeled using a specific protein binding molecule, such as an antibody conjugated to a Feature Barcode oligonucleotide. This protocol provides guidance for antibody-oligonucleotide conjugation and outlines cell surface protein labeling for use with Single Cell RNA sequencing protocols with Feature Barcoding technology. This protocol also provides guidance for enriching labeled cells using Fluorescence Activated Cell Sorting (FACS).

This demonstrated protocol is optimized for counting cells in the range of 700-1200 cells/µl. If using the Single Cell 3' LT v3.1 (low throughput) application, ensure cells are counted as indicated in this protocol and then diluted to the LT specific optimal loading concentration of 100-600 cells/µl using the Cell Dilution Overview in the LT User Guide.

To design a custom conjugate oligonucleotide, use a sequence from the Feature Barcode whitelist (CG000193). Please refer to the following article for details: How do I use the Feature Barcode Whitelist?

FOR USE WITH:

• Chromium Next GEM Single Cell 3' Reagent Kits User Guide (v3.1 Chemistry Dual Index) with Feature Barcoding technology for Cell Surface Protein (CG000317)
• Chromium Next GEM Single Cell 3' Reagent Kits User Guide (v3.1 Chemistry Dual Index) with Feature Barcoding technology for Cell Surface Protein and Cell Multiplexing (CG000390)
• Chromium Next GEM Single Cell 3' Reagent Kits v3.1 User Guide with Feature Barcoding technology for Cell Surface Protein (CG000206)
• Chromium Next GEM Single Cell 5' Reagent Kits User Guide (v2 Chemistry Dual Index) with Feature Barcoding technology for Cell Surface Protein and Immune Receptor Mapping (CG000330)
• Chromium Next GEM Single Cell V(D)J Reagent Kits User Guide v1.1 with Feature Barcoding technology for Cell Surface Protein (CG000208)
• Chromium Single Cell 3' Reagent Kits v3 User Guide with Feature Barcoding technology for Cell Surface Protein (CG000185)
• Chromium Single Cell V(D)J Reagent Kits User Guide with Feature Barcoding technology for Cell Surface Protein (CG000186)
• Chromium Next GEM Single Cell 3ʹ LT Reagent Kits (v3.1 Dual Index) with Feature Barcode technology for Cell Surface Protein (CG000400)