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Highly Sensitive Protein Analysis with Nanogram Inputs

Library Preparation Kit Spincotech
Unlock biological samples with Library Prep Kit V3 for robust library construction from 200 ng protein.

Library Preparation Kits

High-confidence protein sequencing from nanogram inputs with Library Preparation Kit V3.

Protein Sequencing Kits

Achieve deeper protein insights with ready-to-sequence chips, recognizers, and aminopeptidases.

Nanogram input Spincotech

Nanogram Input

Construct NGPS libraries with 200 ng total protein, delivering high-confidence sequencing from limited protein sources

Low-abundance detection Spincotech

Low-Abundance Detection

Inject into mouse model; harvest target tissues

Resolve immunoprecipitates Spincotech

Resolve Immunoprecipitates

Enhanced bioinformatics assess proteins immunoprecipitated from biological samples, like cell lysates, uniquely differentiating proteins in a mixture with precision

Fast and familiar workflow Spincotech

Fast and Familiar Workflow

Updated protocol yields sequenceable libraries within 48 hours, with less than two hours of hands-on time

Reduced Input for Targeted Proteomics

Library Preparation Kit V3 was designed to significantly reduce the input needed to produce high-quality NGPS libraries.
More data from less protein: 5-protein mix libraries were produced from 200 ng compared to 8-40 μg with the V2 kit, delivering elevated peptide coverage with over 95% confidence.
Resolve protein mixtures with confidence: 10-protein mixture sequenced at 200 ng (~20 ng per protein) with the V3 kit compared to 7.6–33 μg (0.76–3.3 μg per protein) from the V2 kit showed improved precision.
Reduced Input for Targeted Proteomics Spincotech

Resolve Immunoprecipitated Protein Mixtures

  • Upgrades to the inference software enhance confidence in identifying proteins in complex mixtures, with only nanogram amounts of input protein.
  • Libraries were generated using the V3 kit from HEK293 cell extracts derived from 106 cells expressing FLAG-tagged integrin alpha-1 (ITGA1) following immunoprecipitation.
  • NGPS accurately identified the ITGA1 protein with top-ranked inference using Sequencing Kit V4 on the Platinum Pro platform.
Resolve Immunoprecipitated Protein Mixtures Spincotech

Sensitive Identification of Protein Mixtures

Proteins have diverse intrinsic properties that impact their performance across proteomics assays. To characterize the limit of detection of low-abundance proteins in a mixture, a panel of five proteins was titrated in mixtures with a constant total protein of 200 ng per library. Here, ADML was maintained at high abundance (40–196 ng), while four others (LMNB1, PD-l1, HSA, and ILI8R) stepped down from 0.5–40% (1–40 ng) per protein to model performance at low abundance.

Libraries were prepared with Library Preparation Kit V3, sequenced on a full chip using Sequencing Kit V4 on Platinum Pro.

Highest-coverage proteins (LMNB1, PD-L1, HSA):
  • Accurately detected with ≥95% confidence
  • LMNB1: reliably inferred at 1 ng in the mixture (0.5% abundance)
  • PD-L1 and HSA: reliably inferred at 2 ng (1% abundance)
Lower-coverage proteins (IL18R, ADML):
  • Required higher mass for high-confidence detection
  • IL18R: required 80 ng (40% abundance) to reach high-confidence inference
  • ADML: required 160 ng (80% abundance)
Sensitive Identification of Protein Mixtures Spincotech