BD® AbSeq Protocols(cell-surface/intracellular protein)

Single Cell Labeling - cell surface protein only

• Single Cell Labelling with BD® AbSeq Ab-Oligos (1 to 40 plex)
  • This protocol describes the use of BD® AbSeq Ab-Oligos (antibody-oligonucleotides) when staining up to 40 antibodies at a time.
  • Doc ID: 23-24262(01)
  • Revision date: 11/2022
• Single-Cell Labeling with BD® AbSeq Ab-Oligos (41 to 100 plex) 
  • This protocol describes the use of BD® AbSeq Ab-Oligos (antibody-oligonucleotides) when staining 41 - 100 antibodies at a time.
  • Doc ID: 23-22314(02)
  • Revision date: 11/2022

Single Cell Labeling - cell surface protein with sample multiplexing(SMK)

• Single Cell Labelling with BD® Single-Cell Multiplexing Kits and BD® AbSeq Ab-Oligos (1 to 40 plex)
  • This protocol describes the use of BD AbSeq Ab-Oligos (antibody-oligonucleotides)(1 plex to 40 plex) with BD® Single-Cell Multiplexing Kits.
  • Doc ID: 23-21339(02)
  • Revision date: 11/2022
• Single-Cell Labeling with BD® Single-Cell Multiplexing Kits and BD® AbSeq Ab-Oligos (41 to 100 plex)
  • This protocol describes the use of BD AbSeq Ab-Oligos (antibody-oligonucleotides)(41 plex to 100 plex) with BD® Single-Cell Multiplexing Kits.
  • Doc ID: 23-22354(02)
  • Revision date: 11/2022
• Single-Cell Labeling with BD® Flex Single-Cell Multiplexing Kits and BD® AbSeq Ab-Oligos (1 plex to 100 plex)
  • This protocol describes the use of BD® AbSeq Ab-Oligos (antibody-oligonucleotides) with the BD® Flex Single-Cell Multiplexing Kits (Cat. no. 633849-633852).
  • Doc ID: 23-24312(01)
  • Revision date: 12/2022

Single Cell Labeling - intracellular protein

• Single-Cell Labeling with BD® AbSeq Ab-Oligos for Intracellular CITE-seq
  • This protocol details how to stain for intracellular antigens using BD® Intracellular AbSeq (IC-AbSeq) antibodies for profiling with the BD Rhapsody™ system. Each BD® IC-AbSeq antibody is conjugated to an antibody-specific oligonucleotide barcode for profiling alongside surface protein and mRNA expression. Note that this protocol should be used alongside BD Rhapsody™ System Single-Cell Labeling with BD® AbSeq Ab-Oligos (1 plex to 40 plex) Protocol, 23-24262 or BD Rhapsody™ System Single-Cell Labeling with the BD® Single-Cell Multiplexing Kit and BD® AbSeq Ab-Oligos (1 plex to 40 plex) Protocol, 23-21339 to support single-cell labeling with BD® AbSeq Ab-Oligos without and with sample multiplexing, respectively. This protocol has been tested for use with up to 40 surface markers.
  • Doc ID: 23-24464(03)
  • Revision date: 4/2024

Co-staining cells with fluorescent antibodies and BD Oligo-Conjugated Antibodies

• • Here, we address how to co-stain cells with the same clone of both fluorescent antibodies and oligonucleotide-conjugated antibodies (BD® Single-Cell Multiplexing Kit or BD® AbSeq Ab-Oligos).
  • Doc ID: 23-23247-00
  • Revision date: 03/2021

These cell labelling protocols are followed by cell capture and then downstream library prep protocols for WTA, Targeted or VDJ(TCR/BCR) analysis.

If you are interested in an AbSeq only readout(without the mRNA readout), follow one of these protocols after cell capture

• With SMK : BD® AbSeq and Sample Tag Library Preparation Protocol
  • This protocol enables users to use cell surface proteins to profile cells independently of mRNA. Included here are instructions for how to generate BD® AbSeq single-cell libraries and Sample Tag libraries using the BD Rhapsody™ Single-Cell Analysis system, or the BD Rhapsody™ Express Single-Cell Analysis system. 
  • Doc ID : 23-24228(01)
  • Revision date : 12/2022
• Without SMK : BD® AbSeq Library Preparation Protocol
  • This protocol enables users to use cell surface proteins to profile cells independently of mRNA. Included here are instructions for how to generate BD® AbSeq single-cell libraries using the BD Rhapsody™ Single-Cell Analysis system, or the BD Rhapsody™ Express Single-Cell Analysis system
  • Doc ID : 23-24227(01)
  • Revision date : 12/2022

Check out data analysis resources specific to Ab-Seq