Make Pepsin Stock Solution. Keep frozen. Dissolve 0.5g of pepsin in 5 ml of distilled water. Concentration of pepsin is 100 mg/ml.
Make Pepsin Working Solution (0.05 mg/ml) Add 20 ul of Pepsin Stock Solution into 40 ml of 10mM HCl. Final concentration of pepsin is 0.05 mg/ml.
1% Formaldehyde Fixative Solution (V=400 ml). Mix 10 ml 37% Formaldehyde with 386 ml PBSx1 and 4 ml 100XMgCl2 100xMgCl2 = 2 M MgCl2. Keep in refrigerator. Discard after each use. Discard solution after each use.
COVERSLIP REMOVAL
Soak slides in Xylene for ~48 h at ambient temperature or until coverslips are easily removed or fall off of the slide.
Wash slides in fresh xylene for 5 min. I would recommend to do the third Xylene wash in fresh solution for 5 min.
Air dry slides for ~15 min
Place slides in coplin jar containing 95% Ethanol. Can be stored for several days/weeks in refrigerator.
Wash slides in coplin jar containing 85% Ethanol, followed by washing in coplin jar containing 70% ethanol, 5 min each wash.
Incubate slides in 2xSSC for 15-60 min. Proceed with pretreatment followed by FISH.
PRETREATMENT PROCEDURE
Incubate slides in 2 X SSC at 37ºC for 10 min.
Incubate slides in Pepsin Working Solution at 37ºC for 13 min.
Incubate in PBS X1 for 5 min, at ambient temperature.
Fix slides in 1% Formaldehyde for 5 min at RT.
Wash slide in PBS X 1 for 5 min at ambient temperature.
Proceed with slide denaturation according to FISH protocol.
Please click "Accept Sale/Sharing and Targeted Advertising" to enable full site functionality.
At this time, we are experiencing problems with broken links on our site. As an interim solution, for full site functionality you must enable functional and advertising cookies. If you continue to opt-out of these cookies, some content on our site may not be viewable.
We use functional cookies to analyze your use of the site, improve performance and provide a better customer experience. We use advertising cookies to allow us, through certain data assigned and obtained from the user's device, to store or share with third parties information related to user's browsing activity in our website, in order to create an advertising profile and place relevant advertising in our website or those third parties websites. For more information about how Abbott uses cookies please see our Cookie Policy and Privacy Policy.
In order to accept functional and advertising cookies, please click "Enable Cookies" and then click "Accept Sale/Sharing and Targeted Advertising" to view the full site.
Collapse
Learn more about cookies
YOU ARE ABOUT TO ENTER AN ABBOTT COUNTRY OR REGION SPECIFIC WEBSITE.
Please be aware that the website you have requested is intended for the residents of a particular country or countries, as noted on that site. As a result, the site may contain information on pharmaceuticals, medical devices and other products or uses of those products that are not approved in other countries or regions.
YOU ARE ABOUT TO ENTER AN ABBOTT COUNTRY OR REGION SPECIFIC WEBSITE.
Please be aware that the website you have requested is intended for the residents of a particular country or countries, as noted on that site. As a result, the site may contain information on pharmaceuticals, medical devices and other products or uses of those products that are not approved in other countries or regions.