Recent advances in digital imaging technology and fluorescent western blotting reagents have resulted in better sensitivity and data quality.
Advantages of fluorescent western blotting:
3 Colors + Total Protein Normalization
Fluorescent western blotting doesn't have to be difficult. Here are a few resources to get you started.
Use this poster to:
Hover over each image for more information.
Seeing high fluorescence background? Try specialized fluorescence blocking buffers, like those with casein. Also consider using LF-PVDF membranes.
Standards too bright? Avoid using standards that contain pink bands as these fluoresce very readily. Try Precision Plus Protein™ All Blue or Precision Plus Protein Unstained Standards instead.
Don't want to see all the marks on your membrane? Mark the membrane with a pencil, not an ink pen.
Bio-Rad's fluorescent western blotting workflow is a seamless integration of products designed to work together to offer guaranteed results. This validated set of solutions will make it easy for you to get better data every time.
Hover over each step in the workflow for more information.
Use the slider to learn more about our fluorescent western blotting image system.
Move beyond traditional laser scanning systems and X-ray film. Get the fluorescence performance you expect with higher multiplexing (three targets vs. two) and more fluorophore options. The performance of film meets the quantitation capabilities of digital.
This is the ChemiDoc™ MP Imaging System.
Why compromise when you can get the best of chemiluminescence and fluorescence in a single imaging system.
Traditional systems offer limited options.
With the ChemiDoc MP System, the superior quantitation of digital meets the sensitivity of
film and delivers the best fluorescence performance with the brightest new fluorophore.
StarBright™ Blue 700 Fluorophore
See up to three signals at once or look at a subset of signals. Plus, you can normalize beyond a single protein with stain-free total protein normalization.
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explore blot combinations.
Explore blot combinations by making a selection on the right.
Teaches as you tap and explore
Picks optimal light source and filters
Suggests exposure times
Bio-Rad presents the StarBright™ Blue 700 Secondary Antibodies, which are fluorescently tagged detection reagents for western blotting. These tagged antibodies are exceptionally bright and stable to photobleaching and offer unparalleled sensitivity and reproducibility in western blotting.
Bio-Rad's hFAB™ Rhodamine Antibodies are fluorescently labeled primary antibodies raised against the housekeeping proteins (HKPs) actin, tubulin, and GAPDH. They are intended to be used for normalizing protein loading in western blotting experiments. The hFAB Rhodamine Antibodies require no secondary antibodies to detect HKPs.
Linker monomer for
StarBright Secondary Antibodies can be used with IR800 antibodies or other common fluorophores to detect multiple proteins on the same blot. Bio-Rad's stain-free technology and/or the hFAB Rhodamine Anti-Housekeeping Antibodies can be used to normalize protein loading.
The StarBright fluorophore (maximum excitation/
emission = 470 nm/700 nm) emits in the far red/near infrared (NIR) range of the spectrum, where there is minimal background.
The exceptional brightness of StarBright Antibodies leads to exposure times orders of magnitude shorter than those of regular fluorescent dyes.
The StarBright fluorophore is conjugated to highly cross-adsorbed antibodies.
Bio-Rad's hFAB Rhodamine Antibodies are fluorescently labeled primary antibodies raised against the actin, tubulin, and GAPDH housekeeping proteins (HKPs).