Invitrogen No-Stain protein labeling reagents provide a flexible, accurate, fast and reliable method for visualization and mapping Dissolve the protein on the gel or membrane (after transfer). It forms covalent bonds with proteins in gels or on membranes within 10 minutes, does not require any destaining steps, and can be visualized immediately using any commonly used imager. No-Stain reagents do not require any specific gel or other reagents and are compatible with gel staining and western blotting workflows.

Continuous visualization of proteins in gels
Coomassie and other gel staining and destaining steps are time-consuming and tedious. No-Stain Protein Labeling Reagent forms covalent bonds with lysine amino acid side chains on all proteins in the gel within 10 minutes. Because lysine is one of the most abundant amino acids, No-Stain reagents detect all proteins on a gel or membrane and emit a strong signal from the covalently bonded reagent with nanogram-level sensitivity.

•Replaces traditional gel staining reagents — Provides more accurate normalization over a wide range of protein lysate concentrations (1–80 µg)
• Sensitive — The lower limit of detection is 20 ng/band
• Specificity — Only binds to the lysine side chain of the protein. Unbound reagents do not emit fluorescence, resulting in excellent signal-to-noise ratio
• Flexible—Convenient stain-free procedures without changing gels. No-Stain reagent requires no staining and can be used with any gel type (precast or hand cast)

Obtain the gold standard for quantitative Western blots
Protein normalization is A critical step in obtaining reliable and reproducible quantitative Western blots. Total protein normalization is considered the gold standard for quantitative Western blotting. Many major journals have developed guidelines for submitting Western blot studies, and relevant citations from the guidelines are shown below.

• “For quantitative comparisons, appropriate reagents, controls, and imaging methods with a linear signal transduction range should be used” – Properties
• "Record how the data were obtained, whether the signal intensity is linearly related to the antigen loading, and how to normalize Uniform protein loading” – Journal of Biological Chemistry
• " Normalize signal intensity to total protein loaded (total protein assessed by staining membrane) if possible” – Journal of Biological Chemistry
• "Housekeeping proteins should not be used without evidence that the procedure does not affect expression" – Journal of Biological Chemistry

An accurate loading control should show signal intensity versus sample load under all experimental conditions linear relationship between them. By labeling the total protein on the membrane with No-Stain reagent, the signal intensity obtained is guaranteed to be linear with the sample load under all experimental conditions (see figure below). Therefore, total protein serves as an ideal loading control when using No-Stain reagent in quantitative Western blot applications.

•Easy-to-use protocol — Label proteins on nitrocellulose or PVDF membranes in 10 minutes
• Uses a variety of imagers with UV or fluorescence light sources Fast Visualization
• Accurate Total Protein Normalization — A wide linear range of protein detection from 1–80 μg enables detection of No-Stain signals as well as signals of target proteins to achieve Accurate total protein normalization
• Sensitive and stable — nanogram-level sensitivity, stable signal compatible with downstream immunoassay steps
• Superior analysis — Housekeeping proteins are susceptible to signal saturation and other biological changes that are not observed when using No-Stain reagent for total protein normalization

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