ChemiDocTM MP Imaging System – Increased Western Blot Quantification Accuracy
Product News Sep 09, 2011
Using the stain-free detection option of the ChemiDoc MP system, researchers can quantify total protein for normalization purposes after transfer to the membrane. This stain-free total protein normalization method produces faster results and overcomes the potential errors associated with normalization using housekeeping proteins.
“I’ve found that using stain-free imaging with the ChemiDoc MP is as good or better than using housekeeping proteins for normalization,” said Aldrin Gomes, assistant professor at the University of California, Davis. “This also saves time and hassle since I don’t need to re-probe the blot for housekeeping proteins.”
Researchers quantifying western blot experiments traditionally normalize data with housekeeping proteins, which often requires stripping and probing the membrane a second time, or probing with multiple antibodies simultaneously. Stripping and reprobing add extra steps to the protocol, consuming reagents, time, and labor. Alternatively, probing with multiple antibodies simultaneously requires reagent optimization over several experiments. Furthermore, published report indicates that normalizing against housekeeping proteins might produce inaccurate results, depending on experimental conditions and sample types.
Assurance at Every Stage of the Western Workflow
The ChemiDoc MP system also integrates with Bio-Rad’s V3 Western Workflow™, a portfolio of Bio-Rad products that shorten the protein separation and blot transfer steps from several hours to 30 minutes. In addition, the V3 Western Workflow offers researchers confidence in their western blot results, providing them a way to visualize their proteins at every stage of the protocol. Using the ChemiDoc MP system, researchers can visualize protein separation on Criterion™ TGX Stain-Free™ precast gels, verify successful transfer with the Trans-Blot®-TurboTM transfer system by imaging the membrane using stain-free detection, and validate the accuracy of western blot results at every step.
Publication-ready Images Made Easy
The ChemiDoc MP system makes it easy for researchers to acquire high-quality images in seconds, whether for chemiluminescence and fluorescence detection or for colorimetric gel and blot documentation. The ChemiDoc MP system remains in focus at any zoom level. This automated, hands-free operation ensures consistent, reproducible, and high-throughput performance.
“The time taken to get a picture with ChemiDoc MP is much faster than other instruments I’ve used,” said Gomes. “Because of the autofocus feature, I can take publication-ready photos within 10 seconds.”
The ChemiDoc MP system delivers superior linearity and comparable limit of detection to film, while delivering quantitative and reproducible data.