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Technical Protocols and FAQs

Explore our multiomics protocols and get answers to your most frequently asked questions.

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Application Notes and eBooks

Application Note: Correlated Expression of Protein and RNA Using Bulk and Single-Cell Proteogenomics

Learn more about the utility of BEN-seq and see how we used the technique to detect 280 proteins and measure the Th1 response on both unstimulated and stimulated cells.

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Application Note: Titration of TotalSeq™ Antibodies in BEN-Seq Reveals Key Expression Differences

We expand on our BEN-seq workflow and demonstrate the robustness of TotalSeq™ antibody staining by showing how biological and technical variables affect protein expression profiles. By titrating antibodies, we develop a low-cost bulk multiomics workflow.

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Application Note: Single-cell Multiomics Reveals Novel Correlations Between Genomic Variants and Protein Expression in AML Patient Samples

Understand how our antibodies were used with the Mission Bio Tapestri platform to reveal novel correlations between genomic variants and protein expression in AML samples.

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Ebook: The Evolution and Future of Single-Cell Proteogenomics

Learn more about applications using TotalSeq™ reagents and how they can contribute to discoveries in precision oncology.

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Application Note: Efficient Multiplexing of Samples Using Hashtags

The addition of TotalSeq™ hashtag antibodies to your CITE-seq experiment allows you to multiplex samples, increase sample throughput, and improve the identification of multiplets.

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TotalSeq™ in Action: Video Protocols and Seminars

Using TotalSeq™ Antibodies for CITE-Seq

Walk through the CITE-seq workflow using TotalSeq antibodies, including cell surface labeling, GEM generation and barcoding, post-GEM cleanup, library amplification, and gene expression, and cell surface protein library construction.

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Overview of the TotalSeq™ Workflow

See how our TotalSeq reagents are designed to seamlessly integrate into the CITE-seq workflow and are compatible with most droplet-based single-cell sequencing methods.

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Introduction to Single Cell Proteogenomics Applications using TotalSeq™ Antibodies

What is proteogenomics? Learn how our TotalSeq reagents enable simultaneous proteomics and transcriptomics analysis in the same cell using the CITE-seq workflow.

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Tech Insights: Titrating TotalSeq Antibodies

Learn tips and tricks for titrating our TotalSeq™️ antibodies, including our recommended methods and the importance of titration for multiomics.

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What Is Cell Hashing?

Learn how our TotalSeq hashtag reagents empower sample multiplexing, provide robust multiplet detection, and minimize batch effects.

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Tech Insights: Spike-In Antibodies to TotalSeq™️ Cocktails

Learn how you can expand our TotalSeq™️ cocktails by spiking-in additional antibodies for your unique research targets.

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Blogs and Technical Notes

BLOG

Single-Cell Multiomics: A Beginner’s Guide to Understanding CITE-Seq and TotalSeq™ Reagents

Multiomics has transformed traditional sequencing experiments and can provide unparalleled protein and genetic analysis in a single experiment. Learn the basics behind techniques like CITE-seq and REAP-seq and discover how our TotalSeq™ reagents integrate into the workflow.

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A Simplified Multiomics Workflow with Pre-Titrated Antibody Cocktails

Understand our process for developing and optimizing our ready-to-use oligo-antibody cocktails and learn how they provide optimal results and reduce costs.

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Tips and Tricks for Titrating TotalSeq™ Antibodies

Titrating TotalSeq™ antibodies for sequencing applications like CITE-seq is critical for optimal results. Read this blog post to understand the importance of titration, alternative titration methods, and the selection of antibody concentrations.

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Overview of Single-Cell Sequencing: Isolation Methods and Techniques

Discover how single-cell sequencing has propelled biology into a new era of discovery and learn about the methods that enable the isolation of single cells.

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Deciphering the Immune Response to COVID-19 with Single-Cell Multiomics

See how researchers are using single-cell multiomics to decipher the immune response after COVID-19 infection.

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A Deeper View of Cancer with Single-Cell Multiomics

Explore recent studies using single-cell multiomics and our TotalSeq™ reagents to transform cancer research and the development of precision therapies.

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Single-Cell Multiomics Reveals the Neuroimmune Landscape

Explore ways researchers are revealing new insights into the function and phenotype of CNS resident immune cells using single-cell multiomics.

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Uncovering Immune Signatures With TotalSeq™ Universal Cocktails

See how researchers are using TotalSeq™ cocktails to uncover new insights about immune pathophysiology during COVID-19 infection.

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Expand Your Single-Cell Genomics Studies with Proteomics

Understand how DNA-barcoded antibodies can be used to couple protein detection and transcriptome profiling and learn how this approach adds value to scientific research.

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New Research Using TotalSeq™ Products

The list below contains selected publications referencing TotalSeq™ products, as well as the original CITE-seq (Stoeckius et al.) and REAP-seq (Peterson et al.) papers. Please note, manuscripts appearing in bioRxiv and medRxiv have not undergone a peer-review process.

TotalSeq-A

An immunophenotype-coupled transcriptomic atlas of human hematopoietic progenitors Zhang X, et al. Nat Immunol. March 2024.
Microfluidics-free single-cell genomics with templated emulsification Clark I, et al. Nat Biotechnol. Nov 2023.
Multiomic single cell sequencing identifies stemlike nature of mixed phenotype acute leukemia and provides novel risk stratification Peretz C, et al. bioRxiv. May 2023.
An inflammatory state remodels the immune microenvironment and improves risk stratification in acute myeloid leukemia Lasry A, et al. Nature Cancer. November 2022.
Single-cell analysis of human non-small cell lung cancer lesions refines tumor classification and patient stratification Leader AM, et al. November 2021.
Joint single-cell measurements of nuclear proteins and RNA in vivo. Chung H, et al. Nature Methods. Oct 2021.
Early IFN-α signatures and persistent dysfunction are distinguishing features of NK cells in severe COVID-19 Krämer B, et al. Immunity. Sept 2021.
Characterizing cellular heterogeneity in chromatin state with scCUT&Tag-pro Zhang B, et al. bioRxiv. Sept 2021.
Scalable, multimodal profiling of chromatin accessibility, gene expression and protein levels in single cells. Mimitou EP, et al. Nat Biotech. June 2021.
Multiplex protein profiling method for extracellular vesicle protein detection Sun Li and Meckes DG. June 2021.
Integrated analysis of multimodal single-cell data. Hao Y, et al. Cell. May 2021.
Simultaneous trimodal single-cell measurement of transcripts, epitopes, and chromatin accessibility using TEA-seq. Swanson E, et al. eLife. April 2021.
Single-cell profiling of myeloid cells in glioblastoma across species and disease stage reveals macrophage competition and specialization. Pombo Antunes AR, et al. Nat Neurosci. March 2021.
Characterizing the molecular regulation of inhibitory immune checkpoints with multimodal single-cell screens. Papalexi E, et al. Nat Genetics. March 2021.
Multi-modal Single-Cell Analysis Reveals Brain Immune Landscape Plasticity during Aging and Gut Microbiota Dysbiosis. Golomb S, et al. Cell. Dec 2020.
Multi-omic single cell analysis resolves novel stromal cell populations in healthy and diseased human tendon. Kendal et al. Scientific Reports. Sept 2020.
Mouse models of neutropenia reveal progenitor stage-specific defects. Muench DE, et al. Nature. June 2020.
T cell-inducing vaccine durably prevents mucosal SHIV infection even with lower neutralizing antibody titers. Arunachalam P, et al. Nat Med. 2020.
Single cell transcriptomics reveals opioid usage evokes widespread suppression of antiviral gene program. Karagiannis et al. Nat Comm. May 2020.
Normalizing and denoising protein expression data from droplet-based single cell profiling. Mulè M, et al. bioRxiv. 2020.
Broad immune activation underlies shared set point signatures for vaccine responsiveness in healthy individuals and disease activity in patients with lupus. Kotliarov et al. Nat Med. Feb 2020.
Single-cell analysis reveals fibroblast heterogeneity and myofibroblasts in systemic sclerosis-associated interstitial lung disease. Valenzi EM, et al. Annals of the Rheumatic Diseases 2019.
Nuclei multiplexing with barcoded antibodies for single-nucleus genomics. Gaublomme et al. Nat Communications. July 2019.
Single-cell multiomic analysis identifies regulatory programs in mixed-phenotype acute leukemia. Granja et al. Nat Biotechnol. Dec 2019.
Multiplexed quantification of proteins and transcripts in single cells. Peterson et al. Nat Biotechnol. Oct 2017.
Simultaneous epitope and transcriptome measurement in single cells. Stoeckius et al. Nat Methods. Sept 2017.

TotalSeq-B

Scalable, multimodal profiling of chromatin accessibility, gene expression and protein levels in single cells. Mimitou EP, et al. Nat Biotech. June 2021.
Single-cell multiomic analysis identifies regulatory programs in mixed-phenotype acute leukemia. Granja et al. Nat Biotechnol. Dec 2019.

TotalSeq-C

Single-cell multiomic analysis identifies regulatory programs in mixed-phenotype acute leukemia. Granja et al. Nat Biotechnol. Dec 2019.
Single-cell multi-omics analysis of the immune response in COVID-19. Stephenson E, et al. Nat Medicine. April 2021.
Maturation and persistence of the anti-SARS-CoV-2 memory B cell response. Sokal A, et al. Cell. March 2021.
Multi-Omics Resolves a Sharp Disease-State Shift between Mild and Moderate COVID-19. Su Y, et al. Cell. Oct 2020
Single-Cell Omics Reveals Dyssynchrony of the Innate and Adapative Immune System in Progressive COVID-19. Unterman A, et al. medRxiv. July 2020
A conserved dendritic-cell regulatory program limits antitumour immunity. Maier B, et al. Nature. 2020.

Resources and Learning

Technical Resources

Titration and Staining

Cell Hashing

Data Analysis

Sample Prep

Technical Protocols and FAQs

Application Notes and eBooks

TotalSeq™ in Action: Video Protocols and Seminars

Using TotalSeq™ Antibodies for CITE-Seq

Overview of the TotalSeq™ Workflow

Introduction to Single Cell Proteogenomics Applications using TotalSeq™ Antibodies

Tech Insights: Titrating TotalSeq Antibodies

What Is Cell Hashing?

Tech Insights: Spike-In Antibodies to TotalSeq™️ Cocktails

Blogs and Technical Notes

Single-Cell Multiomics: A Beginner’s Guide to Understanding CITE-Seq and TotalSeq™ Reagents

New Research Using TotalSeq™ Products

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