TotalSeq oligo-conjugated antibodies have been widely used to add protein data to single-cell RNA and DNA technologies. Now, the same trusted reagents, available in the new TotalSeq-Bn format, can be used to transform the 10x Genomics Visium CytAssist workflow into a multiomic spatial biology assay, with a simultaneous protein and RNA read-out from the same tissue sample.


TotalSeq-Bn antibodies are conjugated to an oligonucleotide containing a sequencing-compatible PCR handle, unique antibody barcode, and sequence complementary to the Visium CytAssist slide oligo pool.




Spatial Proteogenomics Workflow With 10x Genomics Visium CytAssist




The protein and gene expression libraries generated by the Visium CytAssist workflow are shown below. These libraries are analogous to the CITE-Seq libraries generated by the 10x Genomics Chromium platform for single-cell RNA and protein analysis. The main difference is that the 10x Genomics barcode unique for a single GEM is replaced by a spatial barcode that is unique for each of the thousands of spots in the Visium slide. Legacy TotalSeq-B antibodies are not compatible with Visium CytAssist.



Visium CytAssist Spatial Gene Expression – Probe-based Library




Visium CytAssist Spatial Protein Expression Library




TotalSeq™-Bn targets in the Visium Human Immune Cell Profiling Panel


The following TotalSeq™-Bn targets are available as part of the 10x Genomics Human Immune Cell Profiling Panel. Please contact 10x Genomics for information about the Panel and the Visium CytAssist assays.


Protein Marker Description Location Clone Isotype Control
Mouse IgG2a Isotype control Cell surface MG2a-53 N/A
Mouse IgG1, κ Isotype control Cell surface MOPC-21 N/A
Mouse IgG2b, κ Isotype control Cell surface MPC-11 N/A
Rat IgG2a, κ Isotype control Cell surface RTK2758 N/A
PanCK Structural marker Intracellular AE-1/AE-3 Mouse IgG1, κ
PD1 Checkpoint protein Cell surface NAT105 Mouse IgG1, κ
PCNA Cell proliferation marker Nuclear PC10 Mouse IgG2a
BCL2 Apoptotic cell marker Mitochondrial outer membrane 100 Mouse IgG1
Vimentin Mesenchymal cell marker Intracellular O91D3 Mouse IgG2a
CD8a T cell marker Cell surface C8/144b Mouse IgG1, κ
CD45RA Functional T cell marker Cell surface HI100 Mouse IgG2b, κ
CD45RO Functional T cell marker Cell surface UCHL1 Mouse IgG2a
CD19 B cell marker Cell surface R109 Rabbit IgG
PAX5 B cell marker Nuclear 1H9 Rat IgG2a, κ
CD21 Dendritic and B cell marker Cell surface BU32 Mouse IgG1, κ
CD68 Macrophage marker Cell surface BL13756 Mouse IgG1, κ
CD163 Macrophage marker Cell surface QA19A16 Mouse IgG1, κ
HLA-DR Myeloid cell marker Cell surface LN3 Mouse IgG2b, κ
CD66b Neutrophil marker Cell surface 6/40c Mouse IgG1, κ
CD138 Plasma cell marker Cell surface DL-101 Mouse IgG1, κ



TotalSeq™-Bn Single Catalog Products


TotalSeq™-Bn single conjugate antibodies can be supplemented into 10x Genomics Human Immune Cell Profiling Panel. This offers the ability to expand the number of targets that can be interrogated by the Visium CytAssist Spatial Gene and Protein Expression assay. In order to optimize the concentration(s) and pool the additional antibody marker(s), refer to 10x Genomics Custom Add-on Antibody Optimization guide. For more information regarding the TotalSeq™-Bn single antibodies, please reach out to your local BioLegend Field Application Scientist or BioLegend’s Technical Service team.



Compatibility Information  
10x Genomics Visium CytAssist Spatial Gene and Protein Solution with Capture Sequence technology (FFPE) and 10x Genomics Data Analysis Software (Space Ranger and Loupe Browser)
Next-generation sequencing Compatible with Illumina instruments
Tissue FFPE tissue
Slide capture area 6.5 mm2 or 11 mm2



The TotalSeq™-Bn format is not compatible with 10x Genomics Chromium single-cell solutions and CITE-Seq applications. More information about TotalSeq™ in single-cell applications and CITE-Seq can be found here.



Explore Additional Resources


Learn more about the Visium spatial solution:

Related Pages

Remember me
Forgot your password? Reset Password
Request an Account