Automated Jess™ Western Blot Setup with a CyBio FeliX Liquid Handler
Western blotting is a valuable yet laborious method for analyzing proteins, typically involving many steps over 1-2 days. The Jess Simple Western™ automated capillary-based western blot instrument (by ProteinSimple, a Bio-Techne company) greatly streamlines the bulk of the workflow. However, the setup of the Jess plate involves pipetting small volumes of numerous materials; precise pipetting is required to prevent errors. By automating Jess western blot setup with the CyBio FeliX, you can improve workflow efficiency, ensure maximum reproducibility, and free up approximately 60 minutes of hands‑on bench time per run.
Benefits of automating Jess western blot setup with CyBio Felix
Using FeliX for Jess western blot preparation offers several advantages for research and drug discovery:
- High Precision and Accuracy: Felix reliably pipets volumes as small as 1.0 µL, ensuring precise dispensing of expensive reagents.
- Enhanced Reproducibility: Automation greatly reduces user‑to‑user variability, providing data quality that matches or exceeds manual preparation.
- Seamless Software Integration: FeliX uses data from ProteinSimple’s Compass software to identify exactly which antibodies and reagents you need for each specific sample.
- Modular Flexibility: You can easily tailor your protein analysis workflow within the application; for example, adjust antibody combinations or the number of plates prepared per run.
Workflow for preparing a Jess Simple Western plate with CyBio Felix
Deck setup and reagent preparation
FeliX uses data from Protein Simple's Compass software to select the antibodies and reagents to be used with each sample. The Application Studio software guides you through the entire process of deck setup.
Load the deck of the CyBio FeliX with the accessories, reagents, and samples in microfuge tubes, per the screen prompts. Passive cooling adapters are used to maintain stable temperatures for the Jess western blot reagents and cell lysates.
Sample normalization and preparation for denaturization
Based on known protein concentrations, CyBio FeliX dilutes the samples to a standard concentration for optimal Jess western blot analysis. Felix then transfers DTT, master mix, and sample into PCR strip tubes in a 96-well rack.
Once this step is complete, move the samples off deck, seal the tubes, and start the denaturing step in a thermal cycler.
Jess plate preparation (during denaturation)
While samples are denaturing, FeliX loads the Jess plate with the various reagents to perform the analysis: antibodies, buffer, enzyme, substrate, wash buffer, and protein ladder.
The unique layout of the Jess plate is no problem for the Felix liquid handler; it completes the Jess plate preparation in about 22 minutes, so it’s ready by the time that sample denaturation is complete.
Sample transfer to Jess plate
Once the denaturation step is complete, return the samples to the deck and resume the workflow as guided by the Application Studio software.
The FeliX quickly transfers the 24 denatured samples to the Jess plate, completing the plate setup in about three minutes.
Run the plate on the Jess instrument
Upon completion of the FeliX run, remove the Jess plate from the deck, centrifuge it, then place it into the Jess automated western blot instrument.
Jess will take the Simple Western workflow from here!
How to equip your CyBio FeliX for the Jess Application
CyBio FeliX Basic Unit with Enclosure
OL5015-24-100 (1x)
CyBio FeliX Basic unit with enclosure, excluding pipetting head, liquid handling adapters and PC.
CyBio FeliX CHOICE Head
OL3316-14-250 (1x)
For pipetting single wells, columns and rows in 96 and 384 well SBS format. Works with CHOICE adapters.
8 Channel CHOICE Adapter; 1 µl - 50 µl
OL3316-14-332 (1x)
Liquid handling adapter for pipetting columns from 1 µL to 50 µL in 96 well ANSI/SLAS standard format as well as single channel. Requires CHOICE Head.
8 Channel CHOICE Adapter; 10 µl - 1000 µl
OL3316-14-330 (1x)
Liquid handling adapter for pipetting columns from 10 µL to 1000 µL in 96-well ANSI/SLAS standard format as well as single channel. Requires CHOICE Head.
12 Channel CHOICE Adapter; 1 µl - 50 µl
OL3316-14-342 (1x)
Liquid handling adapter for pipetting rows from 1 µl to 50 µl in 96 well ANSI/SLAS standard format as well as single channel; requires CHOICE Head.
Adapter 24 tubes, passive cooling function
844-00136-0 (2x)
Holds up to 24 microfuge or 2 mL tubes with or without attached lids on the FeliX deck; stored chilled, can be used to keep samples cool.
Adapter 96, passive cooling function
844-00135-0 (1x)
Holds a 96 well non-skirted, semi-skirted or full-skirted microplate on the FeliX deck; stored chilled, can be used to keep samples cool.
Microplate Adapter Soft Touch
30-7215-790-24 (1x)
Spring-mounted adapter for microplates within the SBS Format. For optimal transfers of small volumes into empty wells through a slight touch of the well bottom.
Height Adapter 40 mm
844-00445-0
pack of 2
Waste Box & Waste Bags for using with Waste Box
Waste Box for used tips:
844-00430-0
Waste bags for use with waste box
(pack of 50):
10-406-342
TipBox 96/50 µl; PCR-certified, pre-sterilized
30-102-858
TipBox 96/250 µl; PCR-certified, pre-sterilized
30-102-772
Additional materials required
- 12 column reservoir
- 96-well PCR plate, low profile, skirted
- Protein assay kit (BCA or similar)
- ProteinSimple Detection Module
- ProteinSimple 12-230 kDa Separation Module
- See Jess with FeliX application note for a full list of required materials
Related topics and products
Application Note
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