Azure Aqua™ Transfer Cell User Manual
Part Number AC4201
Rev 20200320 Unpacking Instructions
1. Azure Aqua Transfer Cell is packaged in corrugated paper made. Check the contents carefully and please contact us in case of any damage.
2. Contents • Buffer tank and upper lid with connectors • Electrode assembly blotting core • Two transfer cassettes • Six sponges • Two cooling units
3. Please read this user manual carefully. Proper operation of equipment is necessary to avoid damage and to prolong the life of the apparatus.
4. Before using the Azure Aqua Transfer Cell, wash all components with a neutral detergent and rinse well with ultra pure water.
Table of Contents
1. General Overview 2 1.1 General 2 1.2 Technical Specifications 2 1.3 Safety Instructions 2 2. Installation and Basic Operation 3 2.1 Components 3 2.2 Preparation 3 3. Transfer Best Practices 6 3.1 Run Times and Voltage 6 3.2 Best Practices 6 3.3 Procedure for Optimizing Transfer Conditions 6 4. Troubleshooting 8 4.1 During Transfer 8 4.2 After Transfer 8 5. Warranty Policy 8
Azure Aqua Transfer Cell User Manual Page 1 1. General Overview
1.1 General The transfer apparatus contains an easy-to-use lock mechanism in the gel clamp, gel cassettes, and a power plug. A cooling unit is included with the Azure Aqua Transfer Cell. Used frozen, the module absorbs heat generated during electrophoresis, taking the place of an external cooling system.
1.2 Technical specifications
Electrode Assembly outer Polycarbonate Transfer cassette Polycarbonate Electrode Assembly inner Platinum wire Buffer tank and upper lid Polycarbonate Cooling unit Polyethylene and cold storage agent Electrode assembly size 17.5 cm (L) x 13.5 cm (W) x 19 cm (H) Transfer cassette size 10 cm x 11 cm Maximum gel area 7.5 cm x 10 cm Buffer volume (with cooling unit) 650 ml Buffer volume (without cooling unit) 850 ml
Cleaning instructions Wash the electrode assembly, transfer cassettes, and buffer tank with a neutral detergent and warm water. Be careful to avoid damaging the platinum electrodes. Use warm water to rinse the sponges, and then rinse all components with distilled or ultra pure water.
Chemical reagent compatibility Keep all components of the Azure Aqua Transfer Cell away from chloroform, arenes (such as methylbenzene benzene), and acetone. Any damage caused by these reagents is not covered by the warranty policy.
1.3 Safety Instructions Power to the Azure Aqua Transfer cell is supplied by an external power supply. This power supply must be ground isolated in such a way that the DC voltage output floats with respect to ground. ! Regardless of what power supply is used, do not exceed the following input values. Maximum input voltage: 150V (direct current) Maximum input power: 40W Maximum temperature: 40°C
The external power supply provides current to the cell through the lid assembly, providing a safety interlock to the user. When the apparatus is plugged in, a safety mechanism is engaged that disconnects current when the upper lid is lifted and removed from the tank. When using the tank, shut off current prior to removing the upper lid. Do not turn on current before upper lid is in place.
Note: Be sure to follow all safety standards for the proper operation of all Azure products.
Azure Aqua Transfer Cell User Manual Page 2 2. Installation and Basic Operation
2.1 Components
Upper lid
Sponge Filter paper Membrane Gel Filter paper Sponge Transfer cassette
Electrode Cooling unit assembly
Buffer tank
2.2 Preparation Prepare cooling unit by placing into a -20°C freezer. Replace cooling unit in freezer after each use.
1. Prepare the transfer buffer. (Cool to 4°C for optimum heat diffusion.)
2. Prepare components of the transfer stack. For best results, use gloves and forceps when handling all components of the transfer stack. Cut the filter paper and membrane according to the size of gel or use pre-cut filter paper and membranes. Pre-wet the membrane, filter paper, and sponge in transfer buffer (from 15 minutes to 1 hour, depending on the thickness).
Azure Aqua Transfer Cell User Manual Page 3 3. Create the transfer stack. Place the transfer cassette on a clean surface with the black side facing down. Place a sponge on the black side of the cassette. Place the wet filter paper on the sponge, and then place the gel on the filter pater. Next, place the membrane on the gel, followed by more filter paper. Smooth the filter paper over the membrane to remove any bubbles, and add the second sponge.
Sponge Filter paper Membrane Gel Filter paper Sponge Transfer cassette
4. Without moving the gel and filter paper sandwich, close the transfer cassette and lock using the white sliding block.
5. Insert the cassette into the electrode assembly facing the clear side of the transfer cassette towards the red side of the electrode assembly (unless otherwise specified for the assay being performed). Repeat steps 1 through 4 for second gel if necessary.
6. Place the cooling unit in the tank if using and fill the tank with transfer buffer.
Note: Shorter transfer times may not require the use of the cooling unit.
7. Optional: For longer transfers, using a magnetic stir bar and stirring plate can help to keep the temperature within the tank more uniform.
Azure Aqua Transfer Cell User Manual Page 4 8. Place the upper lid over the tank and fit securely into place. Plug the connectors into the power supply and begin transfer. For proper voltage and timing, please refer to Section 3.1
9. After transfer, disassemble the sandwich and carefully remove the membrane using forceps. Rinse the electrode assembly of the transfer cell, the transfer cassette and the sponges well with warm water and rinse with deionized water.
Azure Aqua Transfer Cell User Manual Page 5 3. Transfer Best Practices
3.1 Run times and voltage
Table 3.1 indicates the voltage and time needed for transfer with Azure Transfer Buffer. The general rule is that the shorter the time, the higher the voltage. The cooling unit should be used during overnight transfers.
Buffer Overnight Transfer 1 Hour Transfer Azure Transfer Buffer 12 Volts 55 Volts
Table 3.1 Run times for Azure Transfer Buffer.
The following factors affect resistance and current: • Changing the SDS, acidity, or alkalinity • Thickness of gel • Amount of buffer • Temperature • Time – buffer capacity decreases the longer you transfer
3.2 Best Practices
Equilibrate the gel with transfer buffer Low gel concentrations (<12%) shrink in methyl alcohol solutions, so equilibrating the gel with the transfer buffer can help to achieve cleaner transfers by shrinking the gel prior to transfer.
Use a stir bar Using a stir bar with the Azure Aqua Transfer Cell can help keep buffer conductivity and temperature stable during transfers.
Only adjust pH of transfer buffer if necessary Adjusting the pH can increase conductivity by increasing the current. Adjust as necessary for the proteins being transferred.
3.3 Procedure for optimizing transfer conditions
Optimizing protein transfer
There are several ways to optimize protein transfer: • Use a pH optimized for the proteins being transferred. For most transfers, a pH between 9.5 and 10 is ideal. • Use a non-continuous gradient gel • Limit use of proteases • Maximize contact between the gel and the membrane by smoothing the filter paper to remove bubbles and ensuring transfer sandwiches are very tight.
Azure Aqua Transfer Cell User Manual Page 6 4. Troubleshooting
4.1 During Transfer
If transfer is not performing as expected: • Try increasing the run-time. • Increase the voltage. • Check to make sure that all equipment is assembled properly. • Adjust the pH of your buffer to increase transfer efficiency. If buffer pH is too close to the iso-electric point, it can cause transfer to fail. Be sure to adjust pH accordingly. • If there is protein sediment in the gel, add SDS to the buffer. • Check the power supply. • Decrease the amount of methyl alcohol in the buffer. • Try reducing the gel concentration.
4.2 After Transfer
If the pattern of the gel clamp appears on the membrane: • Lower the voltage of the transfer and transfer for longer at a lower temperature. • Replace the transfer sponges. • Use new transfer buffer.
If the membrane is dry during transfer: • If the membrane is dry, it may look grey or have white marks. Ensure membranes are wet prior to transfer.
High background: • Increase the amount of wash time.
Membrane appears blank after blotting: • Stain the gel and membrane with a total protein stain such as Coomassie blue to determine whether protein was transferred successfully from the gel to the membrane. • Test the reactivity of antibody and antigen by ELISA, radioimmunoassay, double immunodiffusion, or precipitation.
5. Warranty Policy
Standard 1 year warranty. For more information visit https://www.azurebiosystems.com/warranty/
www.azurebiosystems.com • [email protected]
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