PROTEIN DETECTION, QUANTITATION & ANALYSIS

Our One-Step protein gel stains are low-cost, quick, and easy-to-use. Simply add stain to your SDS-PAGE gel without fixing or washing, and stain for 5 to 60 min. These stains are non-toxic for safer handling, and are certified under CCR Title 22 as non-hazardous for drain disposal after pH neutralization. One-Step Blue® replaces Coomassie® for visible staining or near-IR fluorescence. One-Step™ Lumitein replaces SYPRO® Ruby for UV box or gel scanner, and One-Step™ Lumitein UV replaces Oriole™ for UV transilluminator.

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. Proteins can then be visualized on a gel or membrane using a fluorescent gel scanner, allowing detection of as little as 1 ng protein per band. If desired, the prestain can be reversed after scanning, facilitating downstream multi-color western blot analysis. 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.

Thermal shift assay for protein stability

The thermal shift assay, also called Protein Thermal Shift™, differential scanning fluorimetry, or Thermofluor assay, is a simple technique for measuring protein denaturation temperature. It can be used to screen conditions that affect protein thermal stability, such as mutations, ligand binding, and buffer formulations. The assay is rapid and is performed on a quantitative PCR system. Thermal shift is compatible with high throughput screening and requires much less protein than methods like circular dichroism and differential scanning calorimetry.

AccuOrange™ Protein Quantitation Kit is a highly sensitive fluorescence-based assay for quantitating purified protein or antibody samples in 96-well format. The detection range of the assay is 0.1-15 ug/mL protein. AccuOrange™ is much more sensitive than traditional protein quantitation assays such as BCA, Bradford and Lowry, and is more linearity and reproducible compared to the NanoOrange® protein quantitation assay. The assay shows minimal variability between different proteins, and has stable fluorescence signal for up to 16 hours. The AccuOrange™ assay has low tolerance for detergents, and is not recommended for use with cell lysates.

Near-infrared (near-IR) western detection is highly sensitive, and offers advantages of wider linear range and multiplexing capability compared to chemiluminscence detection (see our webinar to learn more). Biotium’s near-IR CF® dyes are the brightest and most photostable available. Learn more about CF®680 and CF®770 dyes for near-IR western, In Cell Western®, and other applications.