SYPRO Red

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Structure of SYPRO Red, if known. m = 5 - 6, n = 2 - 4.

SYPRO Red is a fluorescent dye of the merocyanine type that is used to stain proteins in the course of protein characterization, mostly in SDS-PAGE .

properties

Sypro Red binds like its analogue sypro Orange selectively to proteins than 6.5 kilodaltons (including glycoproteins ) in SDS - micelles , but not to DNA or lipopolysaccharide . The mutagenicity , reproductive toxicity and carcinogenicity of SYPRO Red is unknown. The absorption maxima are at λ = 300 nm and 550 nm, the emission is maximum at λ = 630 nm. The sensitivity of SDS-PA gels with SYPRO Red is around 4–8 ng protein, which is slightly worse than with silver staining . Due to the fluorescence, the SYPRO Red staining has a larger linear dynamic range when estimating the quantity compared to the silver staining or Coomassie , ie a better correlation between the amount of protein and the signal intensity. SYPRO Red is not suitable for staining proteins on Western blots , 2D gels or IEF gels . In contrast to silver and Coomassie staining, acetic acid is used for fixation before staining, but no alcohols such as methanol or ethanol , as SYPRO Red relies on the presence of SDS for staining.

Individual evidence

  1. ↑ a b c Aleksandr Petrovich Demchenko: Advanced Fluorescence Reporters in Chemistry and Biology III: Applications in Sensing and Imaging Volume 3 of Advanced Fluorescence Reporters in Chemistry and Biology . Springer 2011. ISBN 9783642180354 . P. 179ff.
  2. ↑ European patent EP0774122A1. PDF .
  3. ↑ TH Steinberg, LJ Jones, RP Haugland, VL Singer: SYPRO orange and SYPRO red protein gel stains: one-step fluorescent staining of denaturing gels for detection of nanogram levels of protein. In: Analytical biochemistry. Volume 239, Number 2, August 1996, pp. 223-237, ISSN  0003-2697 . doi : 10.1006 / abio.1996.0319 . PMID 8811914 .
  4. ↑ S. Gallagher, D. Chakavarti: staining proteins in gels. In: Journal of visualized experiments: JoVE. Number 17, 2008, S., ISSN  1940-087X . doi : 10.3791 / 760 . PMID 19066521 . PMC 3253607 (free full text).
  5. ↑ a b c d e f data sheet SYPRO Red Protein Gel Stain from Sigma-Aldrich , accessed on November 28, 2013 ( PDF ).
  6. ^ TH Steinberg, RP Haugland, VL Singer: Applications of SYPRO orange and SYPRO red protein gel stains. In: Analytical biochemistry. Volume 239, Number 2, August 1996, pp. 238-245, ISSN  0003-2697 . doi : 10.1006 / abio.1996.0320 . PMID 8811917 .
  7. ^ A b Molecular Probes / Life Technologies: SYPRO Orange and SYPRO Red Protein Gel Stains. 2003. PDF .