View source for Developing Your Immunoblot Membrane

===Materials===
*A flash drive 
*A tube rack
*Sheet protectors
*PICO developing solution 
*15 ml tube
*Ethanol + KimWipes towels

===Method===
# 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''

==Developing Machine==
# 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''
* Copy a paste ponceau scan to your flashdrive
* Make new folder, title with condition and cell type 
* Put all the images from flash drive onto personal computer