Before you start

This topic lists the required hardware, software, AssayMAP cartridges, labware, and reagents for running the On-Cartridge Reaction protocol. If you have questions about these items, contact Agilent Customer Service.

Hardware

Required hardware

 

 

The following figure and table show the components of the AssayMAP Bravo Platform, which is required for running the AssayMAP protocols.

Figure. AssayMAP Bravo Platform components

 

 

Item	Required hardware
1	Gripper upgrade
2	Bravo 96AM Head
3	96AM Wash Station
4	Pump Module 2.0 and two carboys
5	96AM Cartridge & Tip Seating Station
6	Risers, 146 mm
7	Peltier Thermal Station with STC controller
8	Custom plate nest
9	Optional. Thermal plate insert for efficient heat transfer on the Peltier Thermal Station: • Red PCR Plate Insert (shown) • 96 Greiner U-Bottom Thermal Insert • 96 Greiner V-Bottom Thermal Insert • 96 AbGene U-Bottom Thermo Insert
10	Orbital Shaking Station with Control Unit

 

 

 

To avoid a hardware crash and equipment damage if you are using the 25 µL cartridges, ensure that the wash station contains white wide-bore chimneys.

 

Note: The wash station wide-bore chimneys work for both 5-µL and 25-µL cartridges and are standard on wash stations purchased in 2020 onward. The wide-bore chimneys are white plastic, whereas the normal-bore chimneys are a semi-clear plastic. For details, see the 96 Channel Wash Station Maintenance Guide.

Optional equipment

The following equipment is recommended when preparing the samples and reagents:

•	Microplate centrifuge, such as the Agilent Microplate Centrifuge or equivalent

•	Microplate sealer, such as the Agilent PlateLoc Thermal Microplate Sealer or equivalent

•	Microplate reader

 

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Software

 

 

The following table lists the minimum software requirements:

 

Software	Version
Agilent VWorks Automation Control	13.1.5
Agilent Protein Sample Prep Workbench	3.2
Microsoft Excel Required for the reagent volume calculators and method setup tools.	2016

 

 

 

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AssayMAP cartridges

The following table lists the most common AssayMAP cartridges for performing the On‑Cartridge Reaction on the AssayMAP Bravo Platform. But any AssayMAP cartridge can be used with this application. Each cartridge type can be purchased as a rack of 96 cartridges.

Note: This application has not yet been optimized for the 25 µL cartridges.

 

Cartridge type	Agilent part number
AssayMAP Protein A (PA-W) cartridge rack	G5496-60000
AssayMAP Protein G (PG-W) cartridge rack	G5496-60008
AssayMAP Streptavidin (SA-W) cartridge rack	G5496-60010
AssayMAP Resin-Free cartridge rack This cartridge can be used for mock runs or as cartridge placeholders if only partial columns of Protein A, Protein G, or Streptavidin cartridges are required. For details, see How to prepare the cartridges.	G5496-60009

Cartridge use and storage guidelines

See the cartridge box label for storage guidelines.

Follow these guidelines to get the best performance from AssayMAP cartridges:

•	Use only primed and equilibrated cartridges.

Note: The On-Cartridge Reaction app uses cartridges that have been primed in another application. For example, you might run Affinity Purification to immobilize samples on the cartridges that you want to use for On-Cartridge Reaction.

•	Do not allow wetted cartridges to dry out.

Note: Cartridges will not dry out during the course of a normal application run. Cartridges can dry out rapidly if they are exposed to air for extended periods (e.g., >1 hour) after they have been primed and equilibrated.

If you need to store primed and equilibrated cartridges for a short period, ensure that you use the lidded blue rack-receiver plate stack with an appropriate solution in the receiver plate chimneys such that the cartridge tips are submerged in the solution.

See How to store prepared cartridges for guidelines on cartridges that are to be stored for longer than 10 minutes.

•	AssayMAP cartridges are intended to be single-use consumables.

•	The stability of the PA-W, SA-W, and PG-W cartridges after capturing target molecules should be determined empirically.

 

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Labware requirements

Labware requirements vary depending on experimental design. The complete list of labware options and the corresponding deck locations are listed in the following table.

The following figure shows the nine Bravo deck locations for labware.

 

Use only the labware specified for each deck location. Using different labware or placing labware at unapproved deck locations can cause a collision resulting in equipment damage.

 

Figure. Labware locations on the Bravo deck (top view)

 

 

Labware	Manufacturer part number*	Deck location options
12 Column, Low Profile Reservoir, Natural PP	Agilent 201280-100	3—9
8 Row, Low Profile Reservoir, Natural PP	Agilent 201282-100	3—9
Reservoir, Seahorse 201254-100, PP, no walls, pyramid bottoms	Agilent 201254-100	3—9
Reservoir, Axygen Scientific RES-SW96-LP, 86mL pyramid bottom	Axygen Scientific RES-SW96-LP	3—9
96 ABgene 1127, 1mL Deep Well, Square Well, Round Bottom	ABgene AB-1127	3—9
96 Bio-Rad PCR, Hard-Shell, Low-Profile, Full Skirt	Bio-Rad HSP‑9611	3—9
96 Eppendorf 30129300, PCR, Full Skirt, PolyPro	Eppendorf 30129300	3—9
96 Greiner 652270, PCR, Full Skirt, PolyPro	Greiner 652270	3—9
96 Greiner 650201_U-Bottom, Clear PolyPro	Greiner 650201	3—9
96 Greiner 650207_U-Bottom, White PolyPro	Greiner 650207	3—9
96 Greiner 651201_V-Bottom, Clear PolyPro	Greiner 651201	3—9
96 Costar 3363, PP Conical Bottom	Corning Costar 3363	3—9
96 Greiner 675801, Half Area, Flat-Bottom, UV Star	Greiner 675801	3—9
96 V11 Manual Fill Reservoir	Agilent G5498B#049	3—9
96 Red PCR Insert + Eppendorf 30129300, PCR, Full Skirt	Agilent insert (provided) and Eppendorf 30129300	4
96 Red PCR Insert + Bio-Rad PCR, Hard-Shell, Low-Profile, Full Skirt	Agilent insert (provided) and Bio‑Rad HSP‑9611	4
96 Red PCR Insert + Greiner 652270, PCR, Full Skirt, PolyPro	Agilent insert (provided) and Greiner 652270	4
96 Greiner U-Bottom Thermal Insert + Greiner 650201_U-Bottom, Clear PolyPro	Agilent G5498B#126 and Greiner 650201	4
96 Greiner V-Bottom Thermal Insert + Greiner 651201 V-Bottom, Clear PolyPro	Agilent G5498B#126 and Greiner 651201	4
96 AbGene U-Bottom Thermal Insert + AbGene 1127, Square Well, Round Bottom	Agilent G5498B#127 and ABgene AB-1127	4
*For dimensionally equivalent alternatives and other details about the labware, see the Labware Reference Guide in the Literature Library page of the Protein Sample Prep Workbench.

 

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Reagents

The volume, type, and concentration of reagents required for On-Cartridge Reaction vary depending on sample characteristics and the desired analytical result. Consult the Reference library to find literature with specific chemistry examples.

The deck configuration for On-Cartridge Reaction has three to four buffers on the AssayMAP Bravo deck (Equilibration/Chase, Wash 1, Wash 2, and Elution buffer) that are tailored to the sample and desired experimental outcome.

By default, the syringes are rinsed thoroughly with deionized water after completing the protocol to reduce the risk of premature syringe failure due to the buildup of salts within the syringe barrels. To perform more stringent syringe washing between runs, use the Syringe Wash utility. For details, see Syringe Wash v2.1.

All labware requires specific volume excesses for the protocol to execute properly. Use the On-Cartridge Reaction Reagent Volume Calculator to help determine volume requirements for specific protocol conditions. See Preparing the reagents.

 

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