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Stellar Fluorescence Detection Modules

Stellar Detection Modules for Jess – Highest Sensitivity Fluorescence Multiplexing for Simple Western

Stellar™ NIR / IR detection modules for Jess set the industry standard for Western blotting fluorescence detection sensitivity. Stellar fluorescence modules enable detection of low abundance proteins and multiplexing of multiple targets, including multi-color immunoassays alongside total protein staining within the same lane. Use Stellar NIR / IR detection modules with Jess and enjoy the following capabilities:

  • Multiplex multiple targets in the same lane including total / phospho pairs and multiplexed immunoassays alongside total protein normalization
  • Multiplex immunoassays alongside total protein normalization
  • Low picogram protein detection sensitivity using NIR / IR fluorescence channels
  • Perform total protein normalization at low concentration ranges
  • Superior signal linearity across a 4-log dynamic range
  • Full automation of the Western blotting workflow and flexible multiplexing strategies using Jess

 

 

Boost the sensitivity of your fluorescent Western blots with Stellar NIR / IR!

Jess chemiluminescent detection provides low picogram sensitivity for RNase A

Jess Stellar NIR/IR  delivers higher sensitivity for RNase A compared to traditional fluorescent .western blots

 

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  Jess with Chemiluminescence Jess with Steller NIR Jess with Stellar IR Competing Fluorescence Western Blotting Technology
LOD of RNase A 1.1 pg 0.4 pg 0.7 pg > 50 pg

Stellar NIR/IR fluorescence offers comparable sensitivity to Simple Western™ chemiluminescence and higher sensitivity than competing traditional fluorescence western blotting approaches. Comparison of limit of detection for RNase A on Jess using chemiluminescence and Stellar NIR (red bands) and Stellar IR (green bands) detection modules with a leading competitor Western blot fluorescence imaging system. Jess with Stellar fluorescence detected RNase A down to 0.4 pg which is comparable to sensitivity achieved with chemiluminescence detection using Jess (1.1 pg). Linear regression analysis of all Jess results had R2 values >0.99 (see app note for data). Jess with Stellar fluorescence demonstrated 100X greater detection sensitivity than the closest competitive traditional Western blot fluorescence imaging system.

 

HOW DOES IT WORK?

Steller NIR/IR amplifies fluorescent signal in a capillary to deliver high sensitivity.

 

MULTIPLEX MULTIPLE TARGETS IN THE SAME LANE INCLUDING TOTAL / PHOSPHO PAIRS

Stellar NIR / IR modules for Jess enable detection of multiple targets within the same capillary with excellent reproducibility.

Reproducibly multiplex total and phospho AKT in the same capillary with Stellar NIR / IR for Jess.

Multiplex total / phospho protein pairs with Stellar NIR and IR fluorescence detection. Lysates (0.2 mg/mL) from Jurkat cells (treated with calyculin A) and 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. Stellar NIR / IR channels enabled multiplexing total and phosphorylated AKT in the same lane with excellent reproducibility (<10% CVs for pAKT and <5% CVs for Total AKT across 24 lanes).

 

MULTIPLEX MULTIPLE TARGETS ALONG WITH TOTAL PROTEIN NORMALIZATION

With Stellar NIR / IR modules for Jess, not only can you multiplex multiple targets within the same capillary, but you can also perform a Stellar Total Protein Assay and normalize your immunoassay signals for more accurate results.

Use Stellar NIR / IR for Jess to normalize protein with total protein.

Use Stellar NIR / IR for Jess to normalize protein with total protein.

Use Stellar NIR / IR for Jess to normalize protein with total protein.

Normalize multiplexed immunoassay data with the Stellar Total Protein Assay – all in the same sample. Lysates from Jurkat cells either untreated (-) or treated (+) with Calyculin A were run at 0.25mg/mL and 0.15mg/mL (untreated and treated with Calyculin A, respectively) and probed for total AKT (IR, green bands) and phospho AKT (NIR, red bands) using Stellar Mouse IR and Stellar Rabbit NIR Detection Modules. Each antibody was run alone and multiplexed (top, left) in both lysates along with a Stellar Total Protein Assay (top, right). When normalizing to total protein content and using the untreated sample as reference, the peak area values for pAKT and total AKT are adjusted accordingly (bottom), providing a more accurate measurement of the change in protein expression of pAKT (9-fold versus 14-fold) and total AKT (0.8-fold versus 1.2-fold) upon treatment. Stellar NIR / IR provides the ability to multiplex targets while also normalizing to total protein to get even more data out of your sample and accurate quantitation.

 

SPEED UP THE TIME TO DATA FOR YOUR MULTIPLEXING EXPERIMENTS

Stellar NIR / IR modules for Jess reduce your time to data when multiplexing same molecular weight and/or same species antibodies in the same lane as compared to RePlex™ with chemiluminescence or traditional Western blot.

Decrease the time to results with Stellar NIR / IR for Jess when detecting AKT phosphorylation in Jurkat cells treated with Calyculin A.

Decrease the time to results with Stellar NIR / IR for Jess when detecting AKT phosphorylation in Jurkat cells treated with Calyculin A.

Jess offers multiple options for detecting multiple targets including RePlex sequential immunoassays and detection channel-based multiplexing using Stellar IR & NIR fluorescence. Lysates from Jurkat cells either untreated (-) or treated (+) with Calyculin A were run at 0.2 mg/mL and were probed using Mouse Anti-Total AKT and Rabbit Anti-Phospho AKT primary antibodies. Both targets were visualized using Stellar multiplexed fluorescence (Mouse IR and Rabbit NIR) or using RePlex with chemiluminescence (Total AKT in first probe and phospho AKT in the second probe). Both Stellar NIR/IR and RePlex assays detected total AKT and phospho AKT targets within the same lane. Stellar NIR / IR runs were completed in approximately 3 hours compared to the 5-hour run time of RePlex with Jess using the chemiluminescence channel. Both Jess methods dramatically speed up time to multi-target data when compared to traditional Western blotting workflows.

 

Multiplex with the highest fluorescence sensitivity for western workflows using Stellar NIR / IR for Jess!

Talk to your sales representative about ordering Stellar NIR / IR and Stellar Total Protein detection modules for your Jess system today