Developing Your Immunoblot Membrane

Developing your Immunoblot Membrane

After the 1 hour secondary time has passed take your membranes into trays of TBST to wash three times for 10 minutes
Cut new sheet protector for your membranes (they can be in one protector)
Take your pico developing solutions (two bottles on bench: brown and clear) with a 15 ml tube, a pipette, and a tube rack to the scanner
Quickly take your membranes into the plastic (dry a little) and bring to the scanner
AT THE SCANNER mix the brown and white solution in the 15 ml tube and flush
The developing solution is 1:1 so divide the total amount needed by 2

Example: 4.5 ml total so 2.25 ml of brown and 2.25 ml of clear

Open the tray and clean with ethanol
Open the sheet protector and quickly add drops of the developing solution until is covers the membranes
Leave the sheet open and push tray in
On the scanner hit live view to see your membranes and adjust position if needed
Hit application_chemiluminesence_ blot_exposure_ show preview optimal auto exposure
Place red crop box over the area you want captured and hit camera and wait
(optional) manual exposure_ set manual exposure time (ex. 60 sec) _ capture and wait. Make sure you are not moving the membrane during these pictures.
Take a full picture at the end. Application_ colormetric_ exposure (automatically/ faint bands) _ capture
Go to gallery_ take any picture and the one you just took (select both)_ hit merge_ back to gallery
Put all pictures wanted onto the flash drive_ save/send_ save to USB stick
Trash membranes, clean tray with ethanol

Add the ponceau scan to your other developed images