VersaBlot™ Total Protein Normalization Kits are designed for reversible, sensitive and highly linear protein quantification on SDS-PAGE gels and western blot membranes.
- Reversible protein prestain for fluorescent western blot normalization
- Superior linearity for western blot normalization compared to housekeeping proteins
- Highly sensitive and accurate, detect as little as 1 ng protein and 10% difference in protein content
- Fast and easy labeling of proteins or cell lysates with near-IR CF® dyes before SDS-PAGE
- CF®680T (Ex/Em 681/698 nm) for Typhoon™ Cy®5 or IR Short filter or Odyssey® 700 channel
- CF®770T (Ex/Em 764/787 nm) for Typhoon™ IR Long filter or Odyssey® 800 channel
VersaBlot™ Total Protein Normalization Kits are designed for rapid and sensitive protein detection on SDS-PAGE gels and western blot (WB) membranes. The proteins are covalently labeled with near-infrared CF® Dyes so that after electrophoresis, the bands can be directly visualized via fluorescent gel scanning, eliminating the need for any gel staining procedures. The labeled proteins on SDS-PAGE gels can then be transferred to membranes for WB normalization. If desired, the prestain can be reversed after scanning, facilitating downstream multi-color WB using antibodies conjugated to near-infrared CF® Dyes in the same channel (see Protocol Overview below).
For SDS-PAGE gel prestaining, the kits allow detection of low concentration proteins down to 1 ng with minimal background. The dyes do not cause any visible change to the shape or mobility of the bands compared to unlabeled proteins visualized by post-staining methods.
For WB normalization, the kits demonstrate outstanding linearity for quantification of total protein contents over a wide dynamic range (0.1 – 15 ug), outperforming the traditional normalization method based on housekeeping proteins. Compared to total protein post-staining products like the Ponceau S and LI-COR® REVERT™ Total Protein Stain, VersaBlot™ features a prestaining protocol with superior signal-to-noise and sensitivity (able to detect 10% difference in protein content), with comparable reversibility and ease-of-use. The labeling and reversal procedures do not affect protein mobility or detection by antibodies (including phosphoprotein and glycoprotein detection).
Choosing the Right Dye
The VersaBlot™ Total Protein Normalization Kits enable multi-color protein detection on membranes, with or without reversal. It is important to maintain balanced signal intensity across all channels for multi-color imaging (i.e. not having one color dramatically brighter than others). If signal crosstalk is a concern, we recommend to use the VersaBlot™ CF®770T Total Protein Normalization Kit.
See our fluorescent western blotting protocols.
REVERT is a registered trademark of LI-COR Biosciences.