EDC Impressum & DSE

Electrophoresis
Development & Consulting
Dr. Hanspeter Schickle
Wolfgang Gstrein

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Mai 30, 2018

Normally the target molecules of the electrophoretic process must be visualized in a staining process after the separation.
Exceptions are coloured proteins, f.e. Hemoglobins or other dyes themselves used as tracking molecules.
Prestaining (before electrophoresis): DNA-molecules can be tagged, DIGE is a prestaing method before Urea-IEF in a 2D-procedure.
Prelabeling of proteins before SDS using T-Rex.

[Dye-Staining]
[Fluorescence]
[Silver Staining]

Coomassie R 350 & G 250, Coomassie-Violet, Stains All, Indoine Blue

T-Rex, Flamingo, Sypro Ruby, DIGE, CY, Sybr Green, Pro-Q Diamond

AgNO3

Method

Molecule

Dye (company #order number)

Sensitivity*

Info

Dye Staining

proteins

Coomassie R 350 Sigma B4921-20TAB*
Coomassie-Violet 17 Sigma #210579-50g

10-100 ng
50-500 ng

SDS: still the best! Quantifiable
IEF: for lipophylic and Glyco-proteins

DNA

“Stains All” Sigma # E9379-1G
Indoine Blue Sigma #200816-SG (Aldrich)

10 ng
xx

Very easy handling, quantifiable!
In work.....

Fluorescence

dsDNA

dsDNA:GelStar Cambrex #50535
Sybr Green I Molecular Probes #S7563

0.5 ng
0.5 ng

Tagging or staining
Tagging or staining

ssDNA/RNA

Sybr Green II Molecular Probes #S7564

2 ng

Denaturating run: No tagging!

proteins

prelabel: T-Rex Dyeagnostics #T-Rex 310
general: Flamingo: Flamingo #161-0492
phophoryl. Prot.: Pro-Q Diamond #P33301

0.2 -1 ng
0.5 ng
0.5 ng

Linear quantification possible (LQp)
Easy procedure! LQp
Specific reaction! LQp

Silver-Staining

nucleic acids

AgNO3

100 pg

No quantification possible

proteins

AgNO3

300 pg

No quantification possible

* per band, * 0,4 g dye per tablet