![]() This is because they cannot tell you exactly how much of your protein of interest there is, only how much there is relative to another sample.Įven though western blot quantification is only semi-quantitative, you still need to be rigorous in how you quantify your blots to ensure your results are as accurate as possible. Western Blots Are Semi-Quantitativeīefore we take you through the necessary steps for quantifying your western blots, we need to clarify that western blots are only semi-quantitative. This means you can see how the levels of your protein of interest change between time points, conditions, and treatments. The value you obtain is directly proportional to the concentration of your protein of interest.Ī quantitative western blot will allow you to measure relative changes between different conditions, so rather than just saying a protein is present or absent in a sample, you can go further and say your target protein is present x-times higher in one sample compared to another for example. Why Should You Perform Western Blot Quantification?įirst, what do we actually mean by western blot quantification? Put simply, this refers to the measurement of the signal emitted by your target protein. While it can be used to simply show the presence or absence of a protein in a sample, it can tell you so much more if you take the next step and perform western blot quantification.Īs scientists, we love nothing more than quantitative data! Here, we have pulled together the 4 key steps for you to perform western blot quantification. It separates proteins based on size and then uses antibodies to detect specific proteins within the sample. These systems can be upgraded to a broader spectrum (full RGB, far red, and near IR) to enable fluorescence multiplexing with the ChemiDoc to ChemiDoc MP Upgrade Kit ( 12003263).Western blotting is a commonly used lab technique to identify proteins within a sample. Note: Also available are the ChemiDoc Imaging System (12003153), which provides a more limited fluorescence capability than ChemiDoc MP Systems. UV Safety Shield for ChemiDoc MP and ChemiDoc Imaging Systems ( 12003914).Gel Alignment Templates for ChemiDoc MP and ChemiDoc Imaging Systems ( 12003915).Holder for ChemiDoc Sample Trays and UV Shield ( 1708377).Blue Sample Tray for nucleic acid applications with GelGreen or any SYBR Stain ( 12003027).White Sample Tray for Coomassie Blue, copper, silver, and zinc stains ( 12003026).Blot/UV/Stain-Free Sample Tray for Bio-Rad stain-free gels and blots, fluorescence multiplexing, chemiluminescence detection, ethidium bromide and SYPRO Ruby ( 12003028).Image Lab Software to install on your computer.Internal computer with Image Lab Touch Software.with fluorescence, chemiluminescence, or colorimetric detection Blotting - identify and quantify proteins, nucleic acids, protein-ligand interactions, etc.Electrophoresis - protein, including stain-free, and nucleic acid gel imaging and documentation.Performing Stain-Free western blotting using the ChemiDoc MP System provides the ability to rapidly assess electrophoresis and blot transfer quality prior to blot processing, to perform total protein normalization and multiplexing using a wide rance of fluorophores. Installation/Operational Qualification kit available for ensuring installation and calibration is properly executed for highly regulated laboratory environments.Smart Tray Technology automatically recognizes the application-specific trays and adjusts imaging parameters and software options accordingly Sample Trays and Smart Tray Technology - three sample trays available to cover diverse imaging applications.Stain-free protein normalization - stain-free imaging permits the normalization of bands to total protein in both in gels and blots eliminates the need for housekeeping proteins.Ease of use - features include automatic selection of optimal light source by application, auto focus, auto exposure, and preview features ensure optimal images.Multiplex fluorescent western blotting - detect up to three proteins simultaneously and eliminate the need for stripping and reprobing.Improve your imaging capabilities with the ChemiDoc MP - benefit from the advantages of using a wider range of fluorescent reporters (RGB, far red, and near infrared) including StarBright Fluorophores see the fluorophore compatibility chart.All-in-one flexible imaging - get precise, reproducible fluorescence, chemiluminescence, and colorimetric gel and blot detection, analysis and documentation in a single system.Save sample, avoid the errors commonly associated with stripping, reprobing, and cutting blots and answer more complex biological questions by optimizing your experiments to fluorescence. ![]()
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