Bioradiations Staff
Learn how to improve your western blotting results with Bio-Rad’s Quick Tips video series. These 1–2 minute videos show you how to perfect each step of the western blotting process and were designed to accompany the in-depth curriculum available in the Western Blot Learning Center.
Latest videos:
How to Transfer High Molecular Weight Proteins
How to Use Signal Accumulation Mode
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How to Load a Mini-PROTEAN TGX Stain-Free Gel for Electrophoresis
The Mini-PROTEAN Tetra Cell is a vertical mini gel electrophoresis system designed for fast, simple setup. Learn how to correctly load a Mini-PROTEAN TGX Stain-Free Gel in a Mini-PROTEAN Tetra Cell Electrophoresis Chamber. The unique formulation of Stain-Free TGX Gels allows for decreased run times (15 minutes) while maintaining high sample resolution. Apply the techniques in this video to minimize the chance of having a leaky upper buffer chamber in your next set of western blotting experiments.
How to Minimize Band Distortion During Gel Electrophoresis for Western Blotting
Uneven sample migration in polyacrylamide gel electrophoresis (PAGE), also called smiling and frowning, can make it difficult to accurately estimate band size and differentiate between bands on your western blot. Watch this video to learn about common causes of uneven sample migration. These tips will help ensure that you get the best possible resolution in your western blotting experiments.
How to Prepare Mini-PROTEAN TGX Stain-Free Precast Gels for Western Transfer
Watch this video to learn how to safely release your Mini-PROTEAN TGX Stain-Free Gel from the gel cassette. Depending on your downstream application, learn the best technique to prepare your gel for staining, imaging, or blotting.
Choosing Between Nitrocellulose and PVDF Western Blotting Membranes
When it comes to western blotting, choosing the best membrane for your application can mean the difference between crystal clear bands and a blotchy mess. There are numerous factors to consider when selecting the right membrane for your western blotting experiments. Knowing how each parameter effects each result will help you know which one to use. In this video, we explore the four key criteria you should consider when choosing between PVDF or nitrocellulose membranes for your western blotting experiment.
How to Assemble a Trans-Blot Turbo Transfer Pack for Western Blot Transfer
In this video, we show how simple it is to assemble a blotting sandwich and perform western transfer using the Trans-Blot Turbo Transfer System. This technique combines traditional blotting techniques with modern filter paper and buffers, allowing transfer of proteins from gels to membranes in as little as three minutes, with minimal preparation time. The entire system is in a ready-to-use format that allows researchers to obtain their results faster and easier, with reproducibility that is difficult to achieve using traditional blotting methods.
How to Set up a Mini Trans-Blot Cell for Western Blot Transfer
The Mini Trans-Blot Cell provides rapid, high-quality blotting of Mini-PROTEAN Gels. A component of the Mini-PROTEAN Tetra Electrophoresis System, the Mini Trans-Blot Cell accommodates two gel holder cassettes for electrophoretic transfer of mini format gels run in the Mini-PROTEAN Tetra Cell. Watch this video for a step-by-step tutorial on how to properly set up a Mini Trans-Blot Cell blotting sandwich for efficient transfer.
How to Transfer High Molecular Weight Proteins with the Trans-Blot Turbo Transfer System
Many factors affect the transfer of proteins from a gel to a membrane. The gel percentage used during electrophoresis, the way gels are handled prior to transfer, and the membrane type selected can all impact downstream analysis of higher molecular weight proteins. Follow these recommendations for transfer of proteins larger than 150 kDa using the Trans-Blot Turbo Transfer System.
Optimizing the Blocking Step in Western Blotting
The blocking step in western blotting is a critical process. An optimized block can increase your blot sensitivity while drastically reducing background and nonspecific binding. In this video, we explore considerations for a good block and common pitfalls to avoid when selecting a blocking reagent. Keeping these tips and techniques in mind during your next western blotting experiment will help you achieve optimal results the first time.
How to Use EveryBlot Blocking Buffer for Blocking and Antibody Incubation in Western Blotting
Watch this video to learn how to properly dilute EveryBlot Blocking Buffer for faster western blot blocking and antibody incubation. EveryBlot Blocking Buffer is an optimized buffer for membrane blocking prior to immunodetection and for antibody dilution during western blotting. This blocking buffer provides a 5-minute blocking time while maintaining maximum sensitivity for all types of western blots, regardless of detection method.
Selecting a Primary Antibody to Get the Best Results in Western Blotting
Choosing a primary antibody can be overwhelming. Watch this video to learn what to expect and what to look out for when choosing a primary antibody, including how to navigate antibody validation data. Follow these tips when selecting a primary antibody to ensure optimal experimental results.
How to Choose Primary Antibodies for Multiplex Western Blotting
Taking the time to select the best primary antibodies for multiplex western blotting goes a long way toward the success of your experiment. Choosing incompatible antibodies will result in crosstalk or nonspecific binding. To learn best practices for choosing antibodies for your next multiplex western blotting experiment, watch this video.
How to Optimize Primary Antibody Concentration and Incubation for Western Blotting
Once you have chosen the best primary antibody for your western blotting experiment, additional optimization may be necessary to minimize nonspecific binding and background on your blots. Watch this video to explore considerations to keep in mind when establishing the ideal incubation time and dilution of primary antibodies when performing a western blot.
How to Choose Secondary Antibodies for Multiplex Western Blotting
Knowing how to select secondary antibodies for multiplex western blotting is critical for generating optimized data. Watch this video for an in-depth look into best practices and techniques, enabling you to achieve the best possible results.
How to Image a Chemiluminescent Blot
Enhanced chemiluminescence (ECL) is a highly sensitive method for detecting proteins bound to western blotting membranes. Our Clarity family of ECL substrates provides simple, high-performance solutions for all your western blotting needs. In this video, we give a step-by-step tutorial on how to prepare a western blot membrane for chemiluminescent detection.
How to Use Signal Accumulation Mode on a ChemiDoc Imaging System
In this video, we explain how to use Signal Accumulation Mode to image a chemiluminescent western blot on a ChemiDoc Imaging System. Signal Accumulation Mode acquires a series of images to establish the optimal acquisition time and take advantage of the full dynamic range of the imaging system. See how Signal Accumulation Mode mimics traditional film imaging without the need for a darkroom.
Auto-Exposure Settings for Western Blot Imaging in Image Lab Touch Software
Image Lab Touch Software is the intuitive and powerful software package that runs the ChemiDoc, ChemiDoc MP, and GelDoc Go Imaging Systems. Features include secured user accounts, multiplex fluorescence acquisition, autoexposure tools, and multiple data export options. In this video, we highlight the optimal and rapid auto-exposure settings that enable a quick evaluation of your western blot result while taking advantage of the full dynamic range of the imaging system.