5.2.2 Detection

Experimental Procedures
1. Chemiluminescence Detection

(1) Detection with Gel Imaging System
First, place a flat piece of plastic wrap on the tray, then use forceps to lay the membrane flat on the plastic wrap.
Set the appropriate exposure frequency and duration.
Drop the developing solution onto the membrane to ensure it evenly covers the front surface of the membrane.
Perform exposure and development according to the set frequency.
Finally, select the appropriate result image.
(2) Film Development
Take the developing film, film developing clamp, and other tools to the darkroom for operation.
Place a flat piece of plastic wrap on the developing clamp, then use forceps to lay the membrane flat on the plastic wrap.
Drop the developing solution onto the membrane to make it evenly cover the front surface of the membrane, then flatten and seal the plastic wrap.
Place several films neatly on top of the wrapped membrane. Ensure the films are placed in one go and avoid moving them during placement.
Close the developing clamp. The development time can be appropriately shortened or extended according to the expression level of the target protein.
Since multiple films are used, the intensity of results on films at different positions will vary. Finally, select the appropriate result image from the films.
2. Fluorescence Detection

(1)
Drain excess TBST from the membrane; the membrane can be dried. For fluorescent staining, it is essential to ensure the membrane is dry.
(2)
Use a suitable fluorescence scanner, lay the membrane flat, and adjust the instrument to the appropriate excitation wavelength.
Scan and photograph the membrane.
Finally, select the appropriate result image.