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Want to have all technical references by your finger tips? Download our FREE eBooks for WB, IHC, ELISA, Flow cytometry and Molecular biology here. These ebooks contain detailed information regarding the principles, protocols troubleshooting tips and optimization tips for their respective assays. Handy for newbies and veterans alike.
Breast Cancer Regulation
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Weak signal is typically caused by problems in the blocking or washing steps, but can also be caused by a number of other issues. Use these tips to identify and troubleshoot you weak signal results:
Keywords: Weak Signal, Low Signal, Western Blot Troubleshooting
Western blotting separates proteins based on size; large proteins migrate through a polyacrylamide gel matrix slower than small proteins do. However, other factors can influence the migration rate of proteins, resulting in band sizes different than predicted based on protein size alone.
Keywords: Band Size, Wrong Band Size, Western Blot Troubleshooting
Many Western blot problems arise due to issues with the background. Splotches, streaks, or background staining can ruin results. Use these troubleshooting tips to identify and resolve the cause of your background troubles:
Keywords: Flecked Background, Speckled Background, Blotched Background, Western Blot Troubleshooting
High background on a western blot occurs when the background signal of the membrane reduces the signal-to-noise ratio to unreadable levels. Use these troubleshooting tips to identify and resolve the cause of your high background:
Keywords: High Background, Saturated Background, Background Staining, Nonspecific Staining, Western Blot Troubleshooting
Distorted bands can make it very hard to interpret your results. Common distortions include smile shaped bands with the edges trailing upward, diffuse bands that are broad or blurry, and streaked bands that trail off in several directions. Make sure your next blot has even, crisp bands by following these tips:
Keywords: Smile Bands, Distorted Bands, Uneven Bands, Western Blot Troubleshooting
High background in IHC occurs when the contrast between the antibody stain and the background staining is low, resulting in poor data. Use these tips to identify and troubleshoot the cause of your high background:
Keywords: High Background, IHC Troubleshooting
Many conditions affect the outcome of IHC staining. In order to maximize your results, proper identification and troubleshooting of issues is key. Here are some tips to help you improve your IHC staining:
Keywords: Low Signal, Weak Signal, Low Staining, IHC Troubleshooting
Overstaining can result in washed-out or oversaturated results, reducing the sharpness and clarity of a sample. Follow these suggestions for troubleshooting your overstained IHC experiment:
Keywords: Overstaining, Oversaturated Staining, IHC Troubleshooting
A variety of issues can arise during the staining step of IHC, such as non-specific staining. Non-specific staining occurs when the primary antibodies bind to proteins other than the target protein. Here are some tips to reduce non-specific binding when using IHC.
Keywords: Non-Specific Staining, High Background, IHC Troubleshooting
A number of problems commonly arise during ELSIA that can result in low signal and poor color development. Here are some troubleshooting tips to help you improve your experiment and get better data.
Keywords: Low Signal, Faint Color, ELISA Troubleshooting
Matrix Effect occurs when the target antigen interacts with matrix components in plasma or serum samples. These matrix components can be endogenous biological components such as phospholipids, carbohydrates, and metabolites. Matrix components can reduce the binding of the antibody to the target protein, or non-specifically bind the antibody, generating weak or noisy results. Here are some tips to reduce matrix effect:
Keywords: Matrix Effect, ELISA Troubleshooting
ChIP sequencing (ChIP-Seq) is a powerful method for identifying genome-wide DNA binding sites for transcription factors and other proteins. Use this guide to help troubleshoot common issues that arise during ChiP procedures:
Keywords: ChIp, Seq, immunoprecipitation, troubleshooting, tips