Invitrogen™ No-Stain™ Protein Labelling Reagent

Instant Visualization of Proteins in Gels

Coomassie and other gel staining and stain removal steps can be extremely time consuming and laborious. Spotless Protein Labeling Reagent forms covalent bonds with lysine amino acid side chains on all proteins within 10 minutes in a gel. Since lysine is one of the most abundant amino acids, the No-Stain reagent enables the detection of all proteins on a gel or membrane, and the strong signal emission from the covalently bound reagent provides nanogram-level sensitivity.

Total Protein Normalization

Total protein normalization is a useful method for obtaining accurate, quantitative western blotting data, as housekeeping proteins can often be affected by experimental conditions. The Invitrogen No-Stain Protein Labeling Reagent is a fast, easy to use, covalent protein labeling reagent, applied to a membrane after gel transfer that provides sensitive, linear detection of protein for total protein normalization of western blotting data. The Invitrogen No-Stain Reagent can also be used as a fast, sensitive gel stain for visualization of proteins in a gel.








 

No-Stain™ Protein Labelling Reagent

• Flexible use labeling format—Use with any protein gel of choice to perform total protein labeling of the membrane post-transfer. Or use it as a fast protein stain after electrophoresis of gels you do not intend to transfer.
• Easy to use protocol—Mix and incubate with the transferred membrane. The reaction time is 10 minutes. Gel staining is also a 10 minute reaction.
• Flexible visualization—Use a wide-range of imagers with UV, Green LED or Fluorescence (~488 mm) light source, including the iBright FL1500 Imaging System, to capture an image.
• Accurate total protein normalization—Designed to provide accurate total protein normalization over a broad linear range for protein detection: 1–80 μg total protein loaded per well.
• Sensitive and stable signal—Protein bands are detected down to 20 ng and the signal is compatible with downstream antibody detection.

No-Stain™ Comparison of Reagent Linearity with Cleaning Proteins

The graph in Figure below shows the linear signal response versus the amount of protein loaded per well using the No-Stain Protein Labeling Reagent for total protein normalization. Signals from the housekeeping proteins appear to be saturating at higher lysate loads and would provide less accurate normalization of results.

























 

No-Stain™ Protein Labelling Reagent Specifications
Number of reactions	A standard kit will label 40 mini gel-sized membranes or 40 mini gels, or a combination of the two.
Reaction time	10 min.
Gel compatibility	Compatible with any gel type (precast or pour your own) or gel chemistry (Bis-Tris, Tris-glycine, Tris-acetate, Tricine)
Membrane compatibility	PVDF, nitrocellulose
Immunodetection reagent compatibility	Compatible with all downstream immunodetection steps with chemiluminescence or fluorescence-based detection
Excitation and emission maxima	Excitation: ~488 nm (epi blue light), Emission: 590 nm
Detection sensitivity	20 ng per band
Linear range	1 – 80 μg total protein load per lane
Imager requirements	Compatible with a wide range of imagers that have UV or fluorescence light sources, for example, the iBright FL1500 Imaging System
How does it work?	The No-Stain Protein Labeling Reagent forms covalent bonds with a portion of the lysine amino acid side chains on all proteins
Kit Component and storage conditions	No-Stain Activator (800 µL; storage -20⁰C); No-Stain Derivatizer (800 µL; storage -20⁰C); No-Stain Labeling Buffer, 20X (2 x 20 mL each; storage 15-30⁰C)
Shelf-life	1 year from date of manufacture

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