Western blot protocol automation with the ASSIST PLUS pipetting robot and Simple Western™

  • Western blot protocol automation with the ASSIST PLUS pipetting robot and Simple Western™

    Foolproof Jess plate set-up with D-ONE

    Automated western blotting is a cutting-edge technology revolutionizing traditional protein detection and analysis. Employing robotic systems, microscale separation, and advanced imaging technologies streamlines the labor-intensive and time-consuming process of western blotting. Automating the multiple workflow steps increases efficiency, making it an invaluable tool in various fields of biological research.

    Simple Western – developed by ProteinSimple, a Bio-Techne brand – is the only fully automated western blotting solution on the market. The advanced, capillarybased technology enables efficient and accurate high throughput protein separation, detection and quantification, with all assay reagents and samples in 1 plate. Simple Western assays are advancing research and development in many applications, including cancer and immunooncology, cell and gene therapy, regenerative medicine and targeted protein degradation.

    The ASSIST PLUS pipetting robot and D-ONE single channel pipetting module effortlessly fill plates specifically designed for Simple Western instruments – like Jess™ or Abby™ – allowing 100 % hands-free time without worrying about tedious liquid handling. INTEGRA protocols automate the Simple Western plate set-up for chemiluminescence detection of single target, multi-target (using RePlex™) or total protein assays. This application note describes fully automated liquid handling of all 3 assay types by performing chemiluminescence detection and total protein analysis in 1 RePlex normalization assay. The results show equivalent performance to manual plate filling, with comparable on-deck sample and reagent stability.

  • Table of contents

    Automated western blotting is a cutting-edge technology revolutionizing traditional protein detection and analysis. Employing robotic systems, microscale separation, and advanced imaging technologies streamlines the labor-intensive and time-consuming process of western blotting. Automating the multiple workflow steps increases efficiency, making it an invaluable tool in various fields of biological research.

    Simple Western – developed by ProteinSimple, a Bio-Techne brand – is the only fully automated western blotting solution on the market. The advanced, capillarybased technology enables efficient and accurate high throughput protein separation, detection and quantification, with all assay reagents and samples in 1 plate. Simple Western assays are advancing research and development in many applications, including cancer and immunooncology, cell and gene therapy, regenerative medicine and targeted protein degradation.

    The ASSIST PLUS pipetting robot and D-ONE single channel pipetting module effortlessly fill plates specifically designed for Simple Western instruments – like Jess™ or Abby™ – allowing 100 % hands-free time without worrying about tedious liquid handling. INTEGRA protocols automate the Simple Western plate set-up for chemiluminescence detection of single target, multi-target (using RePlex™) or total protein assays. This application note describes fully automated liquid handling of all 3 assay types by performing chemiluminescence detection and total protein analysis in 1 RePlex normalization assay. The results show equivalent performance to manual plate filling, with comparable on-deck sample and reagent stability.

Key benefits

  • Full walk-away western blot protocol automation combines the ASSIST PLUS pipetting robot with the D-ONE single channel pipetting module and the Simple Western Jess. On top of that, the operator benefits from VIALAB's flexibility to create sample preparation protocols for the ASSIST PLUS.
  • Foolproof liquid handling and plate filling with D-ONE's liquid level detection (LLD) and automated GRIPTIPS® pipette tip selection, ensuring precision and accuracy for both low and high volume transfers.
  • VIALAB's labware tool simplifies the labware definition for unique plates, and the D-ONE module can even accommodate irregular well distributions.
  • Simple Western advanced capillary electrophoresis immunoassay technology enables reliable, high throughput, automated western blot analysis of up to 24 samples per run, with results ready in as little as 3 hours.
  • Simple Western chemiluminescence and NIR/IR fluorescence detection provide flexible multiplex analysis capabilities, and ensure high sensitivity when working with precious samples or low abundance targets. RePlex enables 2 immunoassays in a single capillary, and even
    provides total protein detection to accurately normalize protein expression data.
  • The small footprints of the ASSIST PLUS and Jess instruments save space, so they fit easily into any laboratory.

Overview: How to fill the Simple Western Jess plate with the ASSIST PLUS

The ASSIST PLUS pipetting robot and D-ONE single channel pipetting module, together with the Simple Western Jess, automate all the liquid handling steps required to analyze 24 samples, providing a complete walk-away solution for western blot protocol automation.

The whole workflow consists of 2 steps (Figure 1):

  1. Simple Western Jess plate filling with D-ONE and the ASSIST PLUS
  2. Protein analysis with the Simple Western

 

This application note provides Jess plate filling protocols for chemiluminescence detection of single targets, multiple targets (using RePlex) and total protein analysis of prepared samples and reagents for reliable downstream protein analysis using Simple Western Technology.

Downloads: App note and protocols for western blot protocol automation with the ASSIST PLUS pipetting robot and Simple Western™

Illustration of the workflow for western blot protocol automation in 2 steps using the D-ONE single channel pipetting module on the ASSIST PLUS pipetting robot and the ProteinSimple Jess.
Figure 1: 2-step western blot automation protocol.

Plate set-up for RePlex chemiluminescence western blot protocol automation

Experimental set-up:

Deck Position A: Wash buffer (blue)

Deck Position B: B1 – antibody diluent (lavender), streptavidin-HRP (red), luminol-peroxide mix (yellow), RePlex mix (pink); B2 – primary antibody probe 1 (light green-1), secondary antibody probe 1 (green-1), primary antibody probe 2 (light green-2), secondary antibody probe 2 (green-2); B3 – biotinylated ladder (violet); B4-B6 - prepared samples (orange, arrow indicates processing direction)

ASSIST PLUS desk setup Jess
Figure 2: Deck set-up for Jess plate filling to perform RePlex chemiluminescence analysis. Position A: Source – 25 ml reservoir. Position B: Source – INTEGRA tube rack for 1.5/2 ml (B1-B2) and 0.5 ml (B3-B6) microcentrifuge tubes. Position C: Target – Simple Western Jess plate.

RePlex plate set-up

Jess plate set-up for RePlex

Place a 25 ml reservoir on deck Position A and fill it with at least 8 ml of wash buffer. Place the INTEGRA tube rack on Position B with the specific reagent and sample tubes, as indicated in the experimental set-up. As shown in Figure 2, the Jess plate is in landscape orientation on Position C. LLD enables the use of different aliquot sizes, and the D-ONE informs the operator if the liquid volume is insufficient.

Equip the ASSIST PLUS with the 0.5-300 μl D-ONE, and run the VIALAB program 'Jess_plate_setup_RePlex'. The ASSIST PLUS and D-ONE with 12.5 μl long, sterile, filter GRIPTIPS transfer 10 μl of antibody diluent from a 2 ml tube in Position B1 (Figure 2, lavender) to wells B1-B25, C1, E1 and F1 of the Jess plate (Figure 3) in Position C. A 1 μl pre- and post-dispense guarantees precise pipetting while preventing bubble creation during dispensing. By automatically changing GRIPTIPS between different reagents or samples, the D-ONE transfers 10 μl of primary antibody probe 1 from a 1.5 ml microcentrifuge tube in B2 (Figure 2, light green-1) to wells C2-C25 (Figure 3) of the Jess plate in Position C (Figure 4a). The D-ONE transfers 10 μl of streptavidin-HRP/NIR from the second 2 ml screw cap vial in B1 (Figure 2, red) to well D1 (Figure 3). Wells D2 to D25 (Figure 3) are filled with 10 μl secondary antibody probe 1 from a 1.5 ml microcentrifuge tube in B2 (Figure 2, green-1). 10 μl primary antibody probe 2 is transferred from Position B2 (Figure 2, light green-2) to wells E2 to E25 (Figure 3), and 10 μl secondary antibody probe 2 is transferred from B2 (Figure 2, green-2) to wells F2 to F25 (Figure 3).

Using 300 μl sterile, filter GRIPTIPS, the D-ONE transfers 500 μl of wash buffer (Figure 2, blue) from the 25 ml reservoir on Position A to the Jess plate compartments in 2 steps (Figure 4b), as indicated in Figure 3. A slow speed (5) prevents bubble creation during buffer dispensing. Afterwards, 170 μl of luminol-peroxide mix is transferred from the 2 ml tube in B1 (Figure 5, yellow), and 300 μl of RePlex reagent mix from another 2 ml tube in B1 (Figure 5, pink) to the compartments indicated in Figure 3. The D-ONE then transfers the RePlex reagent mix in 2 steps, with a pre- and post-dispense of 10 μl to prevent bubble creation during dispensing.

5 μl of biotinylated ladder is transferred from a 0.5 ml microcentrifuge tube in Position B3 (Figure 2, violet) to well A1 (Figure 3). As indicated by the arrow in Figure 2, the D-ONE transfers 3 μl of prepared sample (Figure 2, orange) from each 0.5 ml microcentrifuge tube in Positions B4 to B6 (Figure 2, orange) to wells A2-A25 (Figure 3). Fast dispensing (speed 10) increases the accuracy for small volumes. The pipette then instructs the user to centrifuge the plate for 5 minutes at 2500 rpm, before proceeding with Simple Western Jess protein separation and immunodetection.

Tips:

  • A prompt can be included in the protocol before starting the sample transfer to instruct the user to prepare the samples during reagent transfer.
  • VIALAB's simplified programming allows plate set-up to be easily adjusted to perform western blot normalization by replacing the second chemiluminescence detection with a total protein analysis, as indicated in the kit insert.
Illustration of how the Simple Western Jess plate is filled with reagents when performing RePlex chemiluminescence analysis of 24 samples.
Figure 3: How to fill the Simple Western Jess plate for RePlex chemiluminescence analysis.
The D-ONE single channel pipetting module on the ASSIST PLUS pipetting robot transfers a) primary antibody and b) wash buffer to the Simple Western Jess plate.
Figure 4: The D-ONE transfers a) the primary antibody and b) the wash buffer to the Simple Western Jess plate.

Plate set-up for chemiluminescence western blot protocol automation

Experimental set-up:

Deck Position A: Wash buffer (blue)

Deck Position B: B1 – antibody diluent (lavender), streptavidin-HRP (red), luminol peroxide mix (yellow); B2 – primary antibody (light green), secondary conjugate (green); B3 – biotinylated ladder (violet); B4-B6 – prepared samples (orange, arrow indicates processing direction)

Graphical representation of the ASSIST PLUS pipetting robot deck set-up with INTEGRA 25 ml reservoir, INTEGRA tube rack, Simple Western Jess plate and D-ONE single channel pipetting module for RePlex chemiluminescence analysis.
Figure 5: Deck set-up for Jess plate filling to perform chemiluminescence analysis. Position A: Source – 25 ml reservoir. Position B: Source – INTEGRA tube rack for 1.5/2 ml (B1-B2) and 0.5 ml (B3-B6) microcentrifuge tubes. Position C: Target – Simple Western Jess plate.

Chemiluminescence plate set-up

Jess plate set-up for chemiluminescence western blot

Prepare a similar deck set-up to the RePlex western blot protocol, but without the RePlex reagent mix or the primary and secondary antibodies for probe 2 (Figure 5).

Select and run the VIALAB program 'Jess_plate_setup_chemiluminescence'. Following a similar procedure as for RePlex chemiluminescence detection, the D-ONE starts by transferring 10 μl of antibody diluent (Figure 5, lavender) to wells B1-B25 and C1 of the Jess plate (Figure 6) in Position C. 10 μl of primary antibody (Figure 5, light green) is then transferred to wells C2-C25, 10 μl of streptavidin-HRP/NIR (Figure 5, red) to well D1, 10 μl of secondary conjugate (Figure 5, green) to wells D2 to D25, and 15 μl of luminol-peroxide mix to all wells of row E, as indicated in Figure 6.

The D-ONE follows this with transfer of the wash buffer (Figure 5, blue), biotinylated ladder (Figure 5, violet) and samples (Figure 5, orange), as described for RePlex chemiluminescence detection, and instructs the operator to centrifuge the plate before proceeding with the automated western blot.

Illustration of how the Simple Western Jess plate is filled with reagents when performing chemiluminescence analysis of 24 samples.
Figure 6: How to fill the Simple Western Jess plate for chemiluminescence analysis.

Plate set-up for total protein western blot protocol automation

Experimental set-up:

Deck Position A: Wash buffer (blue)

Deck Position B: B1 – antibody diluent (lavender), total protein labeling reagent (light green), total protein streptavidin-HRP (green), luminol-peroxide mix (yellow); B3 – biotinylated ladder (violet); B4-B6 – prepared samples (orange, arrow indicates processing direction)

Graphical representation of the ASSIST PLUS pipetting robot deck set-up with INTEGRA 25 ml reservoir, INTEGRA tube rack, Simple Western Jess plate and D-ONE single channel pipetting module chemiluminescence analysis.
Figure 7: Deck set-up for Jess plate filling to perform total protein analysis. Position A: Source – 25 ml reservoir. Position B: Source – INTEGRA tube rack for 1.5/2 ml (B1-B2) and 0.5 ml (B3-B6) microcentrifuge tubes. Position C: Target – Simple Western Jess plate.

Total protein plate set-up

Jess plate set-up for total protein analysis

Set up the ASSIST PLUS deck in a similar way to the chemiluminescence detection protocol, but with the reagents for total protein analysis (Figure 7). 

Select and run the VIALAB program 'Jess_plate_setup_total_protein'. The D-ONE automatically selects GRIPTIPS, and transfers 10 μl of antibody diluent (Figure 7, lavender) into wells B1 and C1 to C25, 8 μl of total protein streptavidin-HRP (Figure 7, green) to row D, 15 μl of luminol-peroxide mix (Figure 7, yellow) to row E, and wash buffer from the reservoir to the compartment of the Jess plate, as indicated in Figure 8. Similar to the western blot protocol for chemiluminescence detection, 5 μl of biotinylated ladder (Figure 7, violet) and 3 μl of each sample (Figure 7, orange) are also transferred to the Jess plate in Position C. The run is completed by the transfer of 10 μl of total protein labeling reagent (Figure 7, light green) into wells B2 to B25, as shown in Figure 8. After finishing the liquid transfers, the instrument instructs the operator to centrifuge the plate, as indicated in the kit.

Illustration of how the Simple Western Jess plate is filled when performing total protein analysis of 24 samples.
Figure 8: How to fill the Simple Western Jess plate for total protein analysis.

Methods and results

Western blotting can be very time consuming and prone to errors. This application note demonstrates a full, walk-away solution for reliable and high throughput automated western blotting, combining INTEGRA's ASSIST PLUS pipetting robot and D-ONE single channel pipetting module with ProteinSimple's Simple Western Jess. 

With RePlex, chemiluminescence and total protein detection were combined in a single assay to prove accurate liquid handling for all reagents when setting up plates with the automated protocols. Reagents, HeLa and C2C12 lysates were prepared according to the protocol in the product insert, with lysate dilutions of 0.64, 0.32, 0.16 and 0.08 mg/ml, together with ready-to-use ERK1 primary and secondary antibodies. Each HeLa or C2C12 dilution was prepared in single tube triplicates as individual samples during plate filling.

Figure 9 shows fully automated western blot normalization of HeLa lysates (lanes 2-13), and C2C12 (lanes 14-25), in triplicate. After ERK1 detection (Figure 9a; left), the primary and secondary antibodies were removed with the RePlex reagent mix, to re-stain samples for total protein analysis (Figure 9a; right). The data sets generated were automatically analyzed, using the Simple Western software tool to visualize the uncorrected (Figure 9b; left) and corrected (Figure 9b; right) ERK1 peak target areas. All 3 replicates of each lysate showed great reproducibility while successfully normalizing ERK1 protein levels in 4 different concentrations, proving accurate liquid handling and confirming on-deck reagent/sample stability during plate set-up.

HeLa lysates (lanes 2-13) and C2C12 (lanes 14-25) were each titrated to 0.64, 0.32, 0.16 and 0.08 mg/ml concentrations, and run in triplicate. a) ERK1 detection (left) and total protein detection (right) and b) automatic quantification of uncorrected and corrected ERK1 peak areas using the Simple Western software.  
Figure 9: Full walk-away western blot normalization with the ASSIST PLUS, D-ONE and Jess using RePlex. HeLa (lanes 2-13) and C2C12 (lanes 14-25) lysates were each titrated to 0.64, 0.32, 0.16 and 0.08 mg/ml concentrations and run in triplicate. a) ERK1 detection (left) and total protein detection (right). b) Automatic quantification of uncorrected and corrected ERK1 peak area using the Simple Western software.

Furthermore, simple fluorescence-based detection was carried out to prove the equivalence in performance between automated and manual plate filling, by removing the luminol-peroxide mix from the VIALAB protocol for chemiluminescence detection (Page 6).

Again, reagents and HeLa lysates were prepared according to the protocol in the product insert, with sample dilutions of 0.64, 0.32, 0.16, 0.08, 0.04 and 0.00 mg/ml (0.1x sample buffer). The primary antibody was prepared by diluting 15 μl of HSP60 and 6 μl of ß-actin in 279 μl of milk-free antibody diluent. The secondary antibody was prepared by diluting 15 μl of anti-rabbit IR and 15 μl of anti-mouse NIR in 270 μl of milk-free antibody diluent.

Figure 10a illustrates the results of automated and manual Jess plate set-up (lane view). Wells 1-13 of rows A to D, as well as the wash buffer compartments, were filled using the D-ONE and ASSIST PLUS. Wells 14-25 of rows A to D were filled manually using a single channel pipette. Both filling methods produced reliable fluorescence data when processing the first 3 dilutions of the HeLa lysates in quadruplicate (0.64 and 0.32 mg/ml) and triplicate (0.16 mg/ml), with CVs within the instrument’s specifications.

Figure 10b illustrates the results when performing fluorescent detection of all 6 HeLa dilutions, by preparing each dilution in 4 single tube replicates on the ASSIST PLUS deck (lane view and electropherogram showing 0.64 mg/ml replicates as representation). Again, CVs met the instrument specifications while showing high detection sensitivity for low abundance targets when automating plate filling with the D-ONE and ASSIST PLUS.

Comparable Jess plate filling is achieved when performing (a) automated (2-13) and  manual (14-25) set-up in one plate with various dilutions of HeLa lysates in quadruplicate (of 0.64 and 0.32 mg/ml) or triplicate (0.16 mg/ml); and (b) fully automated set-up of 24 samples when processing 4 replicates of 0.64, 0.32, 0.16, 0.08, 0.04 and 0.0 mg/ml (0.1x sample buffer) HeLa lysates (lane view and electropherogram showing 0.64 mg/ml replicates as representation).
Figure 10: Fluorescence-based detection results demonstrate comparable Jess plate filling. a) Automated (2-13) and manual (14-25) set-up of a Jess plate with various dilutions of HeLa lysates in quadruplicate (0.64 and 0.32 mg/ml) and triplicate (0.16 mg/ml) (lane view). b) Fully automated set-up of 24 samples when processing 4 replicates of 0.64, 0.32, 0.16, 0.08, 0.04 and 0.0 mg/ml (0.1x sample buffer) HeLa lysates (lane view and electropherogram showing 0.64 mg/ml replicates as representation).

Remarks

  • Labware: The simple labware creation tool in the VIALAB library makes the integration of special plates easier than ever.
  • VIALAB software: VIALAB programs can be easily adapted to your specific pipette, labware and protocols.
  • Partial plates: Programs can be adapted at any time to a different number of samples, giving laboratories total flexibility to meet current and future demands.

Conclusion

  • High throughput western blot protocol automation with the ASSIST PLUS pipetting robot, D-ONE single channel pipetting module and Simple Western Jess eliminates user error and demonstrates CVs below instrument specifications.
  • The sensitivity of your western blots can be increased with Simple Western advanced capillary electrophoresis and immunodetection technology, and accurate and precise plate set-up can be performed with the D-ONE module for ASSIST PLUS.
  • This method can decrease the time needed to run a western blot protocol from days to under 3 hours, by using the ASSIST PLUS and D-ONE to automate Simple Western Jess plate filling.
  • Western blot workflows can be effortlessly accomplished with RePlex, and total protein normalization performed with ease, or multiplexed with simultaneous chemiluminescent and fluorescent assays on the same sample.

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  • Rebecca Bodmer

    Application Specialist

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Instruments and accessories

ASSIST PLUS, Pipetting Robot D-ONE Single Channel Pipetting Module, 0.5-300 µl Tip deck for D-ONE on ASSIST PLUS Rack for 1.5/2 ml microcentrifuge tubes Reservoir Base, 25 ml Multichannel Reagent Reservoir, 25 ml GRIPTIPS®, 12.5 µl LONG, Sterile, Filter (for automation systems) GRIPTIPS®, 300 µl, Sterile, Filter for 96 Channel Pipetting Heads Waste Bags for ASSIST PLUS Bio-Techne: Simple Western Jess System Bio-Techne: HeLa Lysate Controls Novus Biologicals: C2C12 Whole Cell Lysate Bio-Techne: Erk 1 Primary Antibody for Size Assays Novus Biologicals: Beta-Actin Antibody Bio-Techne: HSP60 Antibody Bio-Techne: 12-230 kDa Separation Module Bio-Techne: 12-230 kDa Fluorescence Separation Module Bio-Techne: Anti-Rabbit Detection Module Bio-Techne: Anti-Rabbit IR Detection Module Bio-Techne: Anti-Mouse NIR Detection Module Bio-Techne: Total Protein Detection Module for Chemiluminescence based total protein assays Bio-Techne: RePlex Module

ASSIST PLUS, Pipetting Robot

INTEGRA has developed the ASSIST PLUS pipetting robot to streamline routine pipetting tasks at an affordable price. Using INTEGRA electronic multichannel pipettes, the system:

  • automates pipetting tasks,
  • eliminates physical strain and
  • ensures superior reproducibility and
  • error free pipetting.

Part No. 4505

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ASSIST PLUS Pipetting Robot

D-ONE Single Channel Pipetting Module, 0.5-300 µl

The D-ONE single channel pipetting module offers hands-free transfers from individual tubes or wells on the ASSIST PLUS pipetting robot. This makes it the perfect add-on for labs that want to go beyond the possibilities of multichannel pipetting to access unlimited applications. It is designed to offer easy automation of normalization, hit picking and master mix preparation, freeing up your time while eliminating transcription errors.

Learn more about the D-ONE Single Channel Pipetting Module
D-ONE pipetting module 1ch 0.5-300ul

Tip deck for D-ONE on ASSIST PLUS

  • Supports accommodation of 1 x 12.5 µl/ 125 µl tip box and 1 x 300 µl/ 1250 µl tip box
  • Compatible with D-ONE Pipetting Module 0.5 - 300 µl and 5 - 1250 µl
Learn more about the Tip Deck
D-ONE tip deck

Rack for 1.5/2 ml microcentrifuge tubes

Hold up to 48 tubes on removable carriers for easy loading and storage.

Part No. 4540

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Rack for 1.5/2 ml microcentrifuge tubes

Reservoir Base, 25 ml

Holds disposable reservoir securely during use. One free base is included with every case of bulk or individually wrapped reservoirs.

Part No. 4304

Find out more about Multichannel Reagent Reservoirs
25 ml Reservoir Base

Multichannel Reagent Reservoir, 25 ml

  • SureFlo™ anti-sealing array prevents pipette tip seal off which can cause liquid to pop into the tip, filter or even the pipette itself
  • Unique surface treatment spreads liquid evenly offering even lower dead volumes
  • Reusable base with bold, crisp, clearly visible graduation markings
  • Nest inside each other, making it possible to get twice as many reservoirs in half the space of other products on the market
  • Reducing inventory space requirements and shipping costs
  • Available as crystal clear polystyrene reservoirs or polypropylene reservoirs for improved chemical compatibility
Find out more about Multichannel Reagent Reservoirs
SureFlo™ anti-sealing array

GRIPTIPS®, 12.5 µl LONG, Sterile, Filter (for automation systems)

GRIPTIPS® pipette tips perfectly match the multi-lobe tip fitting, snapping firmly on during loading. This guarantees a perfect seal on every tip, preventing them from loosening, leaking or completely falling off. All tips are precisely aligned horizontally, enabling accurate touch-offs, even when pipetting with 384 tips.

Part No. 6405

Learn more about GRIPTIPS® pipette tips for Benchtop Pipetting Systems
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GRIPTIPS®, 300 µl, Sterile, Filter for 96 Channel Pipetting Heads

Precise and consistent tip seal ensures that all tips on a MINI 96, VIAFLO 96 and VIAFLO 384 are at an identical height for reliable touch offs and consistent pipetting results. An additional tip straightness inspection process ensures all pipette tips access wells even when dispensing to a 384 well plate.

  • For use with 200 and 300µl INTEGRA pipettes and MINI 96, VIAFLO 96 and VIAFLO 384 pipetting heads
  • Tip racks are reinforced to guarantee proper loading of the GRIPTIPS® each and every time

Part No. 6435

Find out more about GRIPTIPS®
Sterile, Filter GripTips for 384 Channel Pipetting Heads

Waste Bags for ASSIST PLUS

Disposable waste bags for the tip waste bin to collect tip waste

Part No. 4570

Find out more about ASSIST PLUS waste bags
Disposable waste bags for the tip waste bin to collect tip waste

Bio-Techne: Simple Western Jess System

Jess automates the protein separation and immunodetection of traditional Western blotting, eliminating many of the tedious, error-prone steps. Just load your samples and reagents into the microplate and Jess separates your proteins by size, and precisely manages antibody additions, incubations, washes and even the detection steps. Come back to fully analyzed results in 3 hours. Go further with high sensitivity multiplexing—her best-in-class fluorescent detection channels and RePlexing capabilities get you all the information you need on your samples in one run. Jess automated western blot system—she's like multiplexed Western blot meets ELISA in one.

Source: Website Bio-Techne

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Bio-Techne: HeLa Lysate Controls

Compatible with Jess™, Abby™, Wes™, Peggy Sue and Sally Sue systems.

Source: Website Bio-Techne

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Novus Biologicals: C2C12 Whole Cell Lysate

C2C12 lysate was prepared by homogenization in modified RIPA buffer (150 mM sodium chloride, 50 mM Tris-HCl, pH 7.4, 1 mM ethylenediaminetetraacetic acid, 1 mM phenylmethylsulfonyl fluoride, 1% Triton X-100, 1% sodium deoxycholic acid, 0.1% sodium dodecylsulfate, 5 ug/ml of aprotinin, 5 ug/ml of leupeptin. Cell debris was removed by centrifugation. Protein concentration was determined with Bio-Rad protein assay. The C2C12 lysate was boiled for 5 min in 1 x SDS sample buffer (50 mM Tris-HCl pH 6.8, 12.5% glycerol, 1% sodium dodecylsulfate, 0.01% bromophenol blue) containing 50 mM DTT.

Source: Website Novus Biologicals

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Bio-Techne: Erk 1 Primary Antibody for Size Assays

Erk 1 Rabbit Primary Antibody, 1X stock (1 mL) for use on Jess™, Abby™, Wes™, Peggy Sue and Sally Sue systems.

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Novus Biologicals: Beta-Actin Antibody

Actin makes up the most abundant protein in many eukaryotic cell types. It polymerizes to form microfilaments that have an array of functions, including regulating contractility, motility, cytokinesis, phagocytosis, adhesion, cell morphology, and providing structural support. At least six types of Actin, falling into three categories, exist in higher eukaryotes. The three types, separated based on isoelectric point, include alpha ( alpha -Skeletal, alpha -Cardiac, alpha -Smooth Muscle), beta ( beta -Non-muscle) and gamma (gamma -Smooth Muscle, gamma -non-muscle) isoforms. Non-muscle beta - and gamma -actin are also known as cytoplasmic actin.

Source: Website Novus Biologicals

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Bio-Techne: HSP60 Antibody

Heat shock proteins (HSPs) are a family of highly conserved stress response proteins. Heat shock proteins function primarily as molecular chaperones by facilitating the folding of other cellular proteins, preventing protein aggregation or targeting improperly folded proteins to specific degradative pathways. HSPs are typically expressed at low levels under normal physiological conditions but are dramatically up-regulated in response to cellular stress. Heat Shock Protein 60 (HSP60), also known as Chaperonin 60 (CPN60), is a mitochondrial matrix protein belonging to a highly conserved family of molecular chaperone and stress response proteins. HSP60 plays a role in stabilizing and refolding proteins in response to heat-shock or other cellular stress.  Full length human HSP60 is 98% identical to mouse and rat HSP60.

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Bio-Techne: 12-230 kDa Separation Module

Compatible with Jess™, Abby™ and Wes™ systems. Includes separation box with Capillary Cartridges, 12-230 kDa Pre-filled Plates (pre-filled with Separation Matrix 2, Stacking Matrix 2, Split Running Buffer 2 and Matrix Removal Buffer), Sample Buffer, Wash Buffer, plus Standard Pack 1 clamshell with DTT, FL Standard master mix with 29 kDa System Control protein. Compatible with Detection module - sold separately. Available sizings include 2x13 capillaries per cartridge (26 data points), 8x13 (104 data points), 2x25 (50 data points) and 8x25 (200 data points).

Source: Website Bio-Techne

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Bio-Techne: 12-230 kDa Fluorescence Separation Module

Compatible with Jess™ systems. Includes separation box with Fluorescent Capillary Cartridges, 12-230kDa Pre-filled Plates (pre-filled with Separation Matrix 2, Stacking Matrix 2, Split Running Buffer 2 and Matrix Removal Buffer), Sample Buffer, Wash Buffer, plus Standard Pack 1 clamshell with DTT, FL Standard master mix with 29kDa System Control protein. Compatible with Detection module - sold separately.

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Bio-Techne: Anti-Rabbit Detection Module

Compatible with Jess™, Abby™, Wes™, Peggy Sue and Sally Sue systems. Module includes: Luminol-S, Peroxide, Antibody Diluent 2, Anti-Rabbit Secondary Antibody, Streptavidin-HRP.

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Bio-Techne: Anti-Rabbit IR Detection Module

Compatible with Jess™ systems ONLY. Module includes: Anti-Rabbit IR Secondary Antibody, Milk-Free Diluent and Streptavidin-NIR. Must be used with Fluorescence Separation Module - sold separately.

Source: Website Bio-Techne

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Bio-Techne: Anti-Mouse NIR Detection Module

Compatible with Jess™ systems ONLY. Module includes: Anti-Mouse NIR Secondary Antibody, Milk-Free Diluent and Streptavidin-NIR. Must be used with Fluorescence Separation Module - sold separately.

Source: Website Bio-Techne

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Bio-Techne: Total Protein Detection Module for Chemiluminescence based total protein assays

Compatible with Jess™, Abby™, Wes™, Peggy Sue and Sally Sue systems. The Total protein detection module for Chemiluminescence based total protein assays includes Luminol-S, Peroxide, Antibody Diluent 2, Total Protein Reconstitution Agent 2, Total Protein Streptavidin-HRP. Compatible with Separation module - sold separately.

Source: Website Bio-Techne

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Bio-Techne: RePlex Module

Compatible with Jess™ and Abby™ systems ONLY. The RePlex Module provides all the reagents needed to remove primary and secondary antibodies in a RePlex assay with two immunoassays or immunoassay with Total Protein assay in a single run.

Source: Website Bio-Techne

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Customer’s voice

From ddPCR, to ELISAs and sample normalization for HPLC, CE-SDS and cell based assay useage, we use the ASSIST in our lab daily. We have 6 across our three labs. They are always reliable, accurate, easy to use, and have a very wide range of applications. In addition to freeing up analysts to perform other work, they have decreased assay variation across multiple assays and users in our labs, for more reliable results with more impactful data! The team at INTEGRA is always willing to help with programming, and is very good at listening to feedback in terms of new accessories and software updates. We are very happy with them!

Regan Sobaje, Capsida Biotherapeutics