SYPRO orange

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Structure of SYPRO Orange, if known. m = 2 - 10, n = 3 - 4.

SYPRO Orange is a fluorescent dye of the merocyanine type that is used in the course of protein characterization for protein staining , usually in an SDS-PAGE .

properties

Sypro Orange binds like its analogue sypro Red selectively to proteins than 6.5 kilodaltons (including glycoproteins ) in SDS - micelles , but not to DNA or lipopolysaccharide . SYPRO Orange is not classified as dangerous according to Regulation_ (EC) _No._1272 / 2008_ (CLP) . The absorption maximum is λ = 470 nm, the emission is maximum at λ = 569 nm. The sensitivity of SDS-PA gels with SYPRO Orange is around 4–8 ng protein, which is somewhat worse than with silver staining . Due to the fluorescence, the SYPRO orange 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 Orange is not suitable for staining proteins on Western blots , 2D gels or IEF gels . In contrast to silver and Coomassie staining, acetic acid is also used for fixation before staining, but no alcohols such as methanol or ethanol , since SYPRO Orange depends 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 data sheet SYPRO Orange 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. ↑ Harvey, PH & Pagel, MD The comparative method in evolutionary biology. Oxford University Press, Oxford (1991).
  8. ^ A b Molecular Probes / Life Technologies: SYPRO Orange and SYPRO Red Protein Gel Stains. 2003. PDF .