What to do if I see empty spots on the Invitrogen Laboratory Equipment membrane?
If you observe empty spots on the membrane, it may be due to air bubbles trapped between the gel and the membrane, preventing protein transfer. Ensure you remove all air bubbles using the Blotting Roller. Another cause could be the use of expired or creased membranes. Always use the iBlot™ 2 Transfer Stacks before the expiration date printed on the package.
Why is my Invitrogen iBlot 2 Laboratory Equipment not transferring proteins to the membrane?
If proteins are not transferring to the membrane, check that the electrical circuit is complete and current is flowing through the device. Also, verify that you are using the correct method. Refer to the manual to ensure the electrical circuit is complete and current is flowing through the device. Be sure to use the correct Method.
What causes high background in Invitrogen iBlot 2 results?
High background can occur if TBST buffers are used for washing. Use PBST or WesternBreeze™ wash solutions instead.
Why is it difficult to close the lid of my Invitrogen iBlot 2?
Difficulty closing the lid can occur if the transfer stack is too thick. The iBlot™ 2 Gel Transfer Device can only support gels of ?1.5mm thickness. Ensure the proper gel is being used. Also, make sure that additional filter pads have not been added and that the stack tray is properly aligned. Only use the supplied filter paper with the iBlot™ 2 Transfer Stacks. Do not remove the transfer stack from the plastic sample tray.
What to do if Invitrogen iBlot 2 Laboratory Equipment signal intensity is similar for different protein loads?
If the signal intensity is similar for different protein loads, it could be due to a high protein load where detection is not within the linear range. It is recommended to decrease the protein load, use more diluted antibody, or perform detection for a shorter time. Optimization may be needed.
What causes protein blow-through in Invitrogen Laboratory Equipment?
Protein blow-through can happen if the transfer time was too long. Reduce the transfer time by 30-second increments. Note that pre-stained markers are charged and tend to blow-through more than regular proteins.
Why are high molecular weight proteins remaining in the gel after transfer with Invitrogen iBlot 2 Laboratory Equipment?
If high molecular weight proteins remain in the gel after transfer, it could be due to an incorrect method or transfer conditions. Use the appropriate method and run time based on the gel type. For mini or midi gels: Perform ethanol equilibration step to improve transfer. Use a Tris-acetate gel to separate the high molecular weight proteins. Increase the transfer time in 30 second increments. For E-PAGE™ gels: Increase the transfer time in 30 second increments. Use Method P3 for 8 minutes.
Why are protein bands distorted on the membrane when using Invitrogen iBlot 2?
Distorted protein bands on the membrane can be caused by a non-uniform electric field around the wells. Ensure the gel is properly flattened using the Blotting Roller. Follow the recommendations to obtain good results.
What causes corrosion of the Top Stack in Invitrogen Laboratory Equipment?
Corrosion of the Top Stack can be caused by incorrect placement. Be sure the Top Stack is placed correctly with the copper electrode facing up. Avoid placing the Top Stack in the inverted position.
What should I do if protein is not binding/transferring to membrane (PVDF) in Invitrogen iBlot 2?
If protein is not binding/transferring to a PVDF membrane, it may be because the membrane was dry or partially dry. Regions where PVDF membranes are dry appear whiter than places where the membrane is wet. Remove the membrane, reactivate it in 100% methanol, and rinse in water before reapplying to the transfer stack.
Details about the components included with the iBlot™ 2 Gel Transfer Device.
Safety information and precautions to take before using the product.
Instructions and conditions for setting up the iBlot™ 2 Gel Transfer Device.
Procedures for safely unpacking the instrument upon receipt.
Overview of the front view of the iBlot™ 2 Gel Transfer Device.
General explanation of the system, its features, components, and operation.
Detailed descriptions of the individual components of the device.
Covers setting date/time, tutorials, logs, firmware, factory reset, methods, and parameters.
Technical specifications for the iBlot™ 2 Gel Transfer Device and stacks.
Information on compatible accessories and related products.
Outline of the steps for performing western blotting and general recommendations.
Lists compatible gel types for use with the iBlot™ 2 system.
Step-by-step instructions for assembling the device and performing blotting.
How to choose an appropriate blotting method on the device interface.
Procedure for carefully removing the gel from the cassette.
Detailed steps for correctly assembling the transfer stack components.
Instructions on how to conduct the actual protein transfer process.
Steps for taking apart the transfer stack after the blotting procedure.
Steps to create and program new blotting methods from scratch.
Procedure for naming and saving newly created custom methods.
Guide on using pre-set templates or saved methods as a starting point.
How to modify voltage and time settings for individual steps in a method.
Instructions for adding or deleting steps within a custom method.
Overview of common issues and how to resolve them.
Troubleshooting steps when the device does not detect current.
Solutions for problems like no protein transfer or empty spots on the membrane.
Addressing issues with distorted protein bands or protein blow-through.
Troubleshooting problems with protein binding, high background, and membrane discoloration.
Procedures after blotting, including immunodetection, storage, and staining.
Strategies for improving protein transfer efficiency through parameter adjustments.
Instructions for cleaning and routine upkeep of the instrument.
Procedure for replacing the device's fuse.
Step-by-step guide for replacing worn electrical contacts.
Detailed technical specifications for the main device.
Detailed technical specifications for the transfer stacks.
Ordering information for iBlot™ 2 Transfer Stacks.
Lists other available reagents, precast gels, and premade buffers.
General warnings and precautions for instrument operation, including physical and electrical hazards.
Safety precautions related to electrical hazards and power supply.
Safe procedures for cleaning and decontaminating the instrument.
Guidelines for safely handling chemicals and biological materials.
Explains instrument symbols and lists relevant safety and EMC standards.
Information on how to contact support for the product.
Details regarding the product's warranty policy.
General| Voltage Range | 20-25 V |
|---|---|
| Technology | Dry blotting technology |
| Application | Protein Transfer |
| Power Supply | 100-240 V AC |
| Transfer Time | 7 minutes |
| Compatible Membranes | PVDF, Nitrocellulose |
| Gel Size | Mini and midi gels |
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