The AccuOrange™ assay is a fluorescent dye-based assay. The dye binds to proteins primarily through hydrophobic interactions. Proteins denature upon heating; the dye binds to the exposed hydrophobic pockets of the protein after cooling. The free AccuOrange™ dye is fluorogenic due to non-radioactive decay but becomes highly fluorescent due to the rigid conformation inside the pocket.
The AccuOrange™ assay more sensitive than traditional protein quantitation assays such as BCA, Bradford and Lowry, and shows superior linearity and reproducibility than the NanoOrange® protein quantitation assay (Thermo Fisher Sci.), but has low tolerance for detergents like SDS and Triton® X-100.
We do not recommend staining proteins on the polyacrylamide gel with the One-Step protein gel stains before western blotting as it can significantly reduce protein transfer to the membrane. If protein detection on the gel prior to transfer is desired, the Total Protein Prestains would be more suitable.
The VersaBlot™ Total Protein Normalization Kits allow simple, sensitive and highly linear protein quantitation on SDS-PAGE gels and western blot membranes. The kits allow you to label purified proteins or cell lysates with our near-infrared CF® dyes before running the samples on SDS-PAGE. After electrophoresis, the bands can be visualized on the gel using a fluorescent gel scanner, allowing detection of as little as 1 ng protein per band. Labeled proteins also can be transferred to membranes for western blotting. The staining demonstrates excellent linearity for quantitation of total protein over a wide dynamic range, outperforming traditional western blot normalization based on housekeeping protein detection and staining is reversible.
One-Step Lumitein™ Protein Gel Stain is a ready-to-use luminescent red protein gel stain. It is a dramatically improved version of our original Lumitein™ protein gel stain in terms of convenience and safety. One-Step Lumitein™ can detect as little as approximately 1-10 ng of protein per band.
One-step staining procedure, no fixation required
Non-toxic formulation is safe for easy disposal
Sensitive protein detection down to 1-10 ng
Image with a UV transilluminator, a blue light transilluminator, or laser-based gel scanner
1 L and 4 L sizes
Store at 4°C
Fast and simple procedure
One-Step Lumitein™ gel staining requires only a single 5-60 minute staining step without fixation. Destaining is optional. One-Step Lumitein™ can detect as little as approximately 1-10 ng of protein per band depending on the staining method used, although staining intensity varies between proteins. Staining is fully compatible with mass spectrometry and Edman-based sequencing. Staining can be imaged using a UV transilluminator with EtBr or SYBR filter set, a blue light transilluminator, or a laser-based gel scanner.
One-Step Lumitein™ is non-toxic
One-Step Lumitein™ Protein Gel Stain offers safer handling and ease of disposal compared to other protein gel stains, because it is an aqueous-based solution that does not contain hazardous methanol or acetic acid. One-Step stains are certified under CCR Title 22 as non-toxic to the environment for drain disposal after a simple pH neutralization step. Furthermore, One-Step Lumitein™ costs significantly less than other fluorescent protein gel stains. One-Step Lumitein™ Protein Gel Stain is available in 1 liter bottle or 4 liter Cubitainer® sizes.
One-Step Blue is a registered trademark of Biotium. Lumitein and its related technologies are covered by US and international patents. SYPRO is a registered trademark of Molecular Probes, Inc. Oriole and Flamingo are trademarks of Bio-Rad Laboratories. Cubitainer is a registered trademark of Hedwin Corporation.