EDC Impressum & DSE

Electrophoresis
Development & Consulting
Dr. Hanspeter Schickle
Wolfgang Gstrein

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Visualization: Immuno-Fluorescence

TRex310

conjugated to Goat anti-human IgG

Mai 30, 2018

Due to a new dye (T-REX 310) - conjugated to the anti-human IgG antibody  - the immunofluorescence staining methods getting possible with the same gel used also for silver-staining.
After 2 hours reaction with this Goat anti-human IgG (H+L) / T-Rex 310  and 3 hours washing out of the exceed conjugate, the gel goes directly to a Fluorescence-Imager.
The ReDye physical properties: 648 nm excitation, 663 nm emission.

Advantage: Quick, easy, sensitive and IgG-selective staining.
Disadvantage: Additional hardware is necessary.

Tested instruments:

Fujifilm: FLA 7000 and FLA 5100:
Settings: red laser, 40 um resolution, amplification 825 pmt, filter 630 nm

See the IEFGel 6-11 manual on “Downloads”, page 8.

Rehydration Pool: immunoreaction
pfeil90

The immunoreaction is done in the adequate rehydration pool in 30 ml (300 ul Antibody), 2 hours slightly and horizontally rocked.

pfeil225
steel tray: washing out exceed dye

1 hour: Washing out exceed amount of the antibody in the adequate staining tray

immunfluorescence

Gel: IEFGel 6-11 40S, Samples: E.Dubois Neurologie Tübingen (Prof.Melms)
Visualization: Immuno-Fluorescence with a special conjugated antibody