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Stellar™ Sets the Industry Standard for Western Blotting Fluorescence Detection Sensitivity

Stellar on Simple Western's Jess is a multiplex fluorescence immunoassay and chemiluminescence immunoassay

  • Multiplex low abundance targets including total and phospho-protein isoforms alongside total protein normalization
  • Low picogram protein NIR/IR detection sensitivity
  • Simultaneous total protein normalization at low concentration ranges
  • Superior signal linearity across a 4-log dynamic range
  • Full automation of Western blot analysis plus flexible multiplex strategies
Industry-leading NIR/IR fluorescence detection sensitivity with Stellar

Stellar Boosts Fluorescence Detection Sensitivity for Your Fluorescence and Chemiluminescence Immunoassay

Stellar fluorescence detection is 100X more sensitive than traditional fluorescent western blots

  • Stellar NIR/IR fluorescence detection offers comparable sensitivity to Simple Western™ chemiluminescence and higher sensitivity than competing traditional fluorescence western blotting approaches. 
  • Stellar fluorescence immunoassay demonstrated 100X greater detection sensitivity than the closest competitive traditional Western blot fluorescence imaging system.

Right Figure and Table Below: Comparison of the limit of detection for RNase A on Jess using chemiluminescence immunoassay and Stellar NIR (red bands) and Stellar IR (green bands) detection modules with a leading competitor Western blot fluorescence imaging system.

Chemiluminescence Immunoassay and Stellar Fluorescence Immunoassay LOD RNase A
  Chemiluminescence Stellar NIR Stellar IR Traditional Fluorescence Western Blot
LOD of RNase A 1.1 pg 0.4 pg 0.7 pg > 50 pg

How Does Stellar Work?

Stellar Fluorescence Immunoassay Schematic

Multiplex Fluorescence and Chemiluminescence Immunoassays with Total Protein Normalization

Accurate Protein Expression Measurements, Including Total and Phospho-Protein Isoforms

Multiplex multiple targets and perform the Stellar Total Protein Assay to normalize your fluorescence immunoassay signals for more accurate results – all in the same sample.

Right Figure: Lysates from Jurkat cells either untreated (-) or treated (+) with Calyculin A were probed for total AKT (IR, green bands) and phospho AKT (NIR, red bands). Each antibody was run alone and multiplexed (left panel) in both lysates along with a Stellar Total Protein Assay (right panel).

Note: Stellar Total Protein Detection Module requires the use of a Stellar NIR and/or IR Detection Module in the same run.

Stellar Fluorescence Immunoassay and Total Protein Assay


Right Figure: The peak area values for pAKT and total AKT are corrected using total protein providing an accurate measurement of protein expression.

Stellar Total Protein Correction

Multiplex Fluorescence and Chemiluminescence Immunoassays with Excellent Reproducibility

Stellar NIR/IR Detection channels enabled multiplexing total and phosphorylated AKT in the same lane with excellent reproducibility.

Right Figure: Jurkat whole-cell lysate probed using mouse anti-total AKT and rabbit anti-phospho AKT primary antibodies and Stellar Mouse IR (green bands) and Stellar Rabbit NIR (red bands) Detection Modules. <10% CVs for pAKT and <5% CVs for Total AKT across 24 lanes

Stellar NIR/IR Detection Channel Reproducibility

Multiplex with High Efficiency and Get Stellar Results in 3 Hours

Dramatically Reduce Time to Results Compared to Traditional Western Blotting Workflows


Stellar NIR/IR detection modules reduce your time to data compared to RePlex™ and traditional Western blot when multiplexing with antibodies of the same species and/or antibody targets of similar size

  • Both Stellar NIR/IR and RePlex assays detected total-AKT and phospho-AKT targets within the same lane.
  • Stellar NIR/IR detection runs were completed in approximately 3 hours compared to the 5-hour run time of RePlex
  • Both Stellar and RePlex dramatically speed up time to multi-target data when compared to traditional Western blotting workflows
Stellar Versus RePlex

Order Stellar Fluorescence Detection Modules

Stellar Detection Modules must be used with Fluorescence Separation Modules. The Stellar Total Protein Detection Module must be used with a Stellar NIR/IR Detection Module in the same run. Standard Chemiluminescence Detection Modules may be used with Stellar Fluorescence Detection Modules. See our Simple Western Multiplex Assay Compatibility Chart.

Module Part Number
12-230 kDa Fluorescence Separation Module Order SM-FL001
66-440 kDa Fluorescence Separation Module Order SM-FL002
2-40 kDa Fluorescence Separation Module Order SM-FL003
Anti-Rabbit Stellar IR Fluorescence Detection Module Order DM-013
Anti-Rabbit Stellar NIR Fluorescence Detection Module Order DM-014
Anti-Mouse Stellar IR Fluorescence Detection Module Order DM-015
Anti-Mouse Stellar NIR Fluorescence Detection Module Order DM-016
Stellar Total Protein Fluorescence Detection Module Order DM-TP03
Stellar Training Kit Order PS-T007
Stellar Fluorescence Detection Module Product Foreground