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methods:sop:zipglua23y

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methods:sop:zipglua23y [2023/07/14 13:54] – ohardtmethods:sop:zipglua23y [2023/07/15 15:20] (current) – external edit 127.0.0.1
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 ===== TRIS-saline, pH 7.00 solution ===== ===== TRIS-saline, pH 7.00 solution =====
-Add 1mL of Tris 1M (pH 7.00) to 9 mL sterile saline. Mix well. +Add 1 mL of Tris 1 M (pH 7.00) to 9 mL sterile saline. Mix well. 
  
  
 ===== ZIP 10 nmol/µl ===== ===== ZIP 10 nmol/µl =====
-  - Add 104 µl of Tris-saline to a bottle containing 2 mg of ZIP (ZIP is in the -20 freezer in bottles containing 2mg of lyophilized powder).+  - Add 104 µl of Tris-saline to a bottle containing 2 mg of ZIP (ZIP is in the -20 freezer in bottles containing 2 mg of lyophilized powder).
   - With the same pipette, pipette up and down to dissolve all ZIP, if necessary pass some of the solution by the wall to make sure there is no powder lefts on the walls. If bubbles are formed you can remove them by breaking them with a needle. Put the tube on ice.   - With the same pipette, pipette up and down to dissolve all ZIP, if necessary pass some of the solution by the wall to make sure there is no powder lefts on the walls. If bubbles are formed you can remove them by breaking them with a needle. Put the tube on ice.
   - Take the amount necessary for one experiment (at least 20 µl), put in a small tube and aliquot the rest in at least 20 µl and put in the -20 freezer.    - Take the amount necessary for one experiment (at least 20 µl), put in a small tube and aliquot the rest in at least 20 µl and put in the -20 freezer. 
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-===== GluR23Y peptide ===== +===== GluA2-3Y peptide ===== 
-  - Stock solution +  - **Stock solution** 
-    -  Add 126 µl of Tris-saline to a tube containing 1 mg of GluR23Y (GluR23Y is in the -20 freezer in small tubes containing 0.9 to 2 mg, you have to adjust the amount of Tris-saline according to the amount is written in the tube). Mix well and place the tube on ice.+    -  Add 126 µl of Tris-saline to a tube containing 1 mg of GluA2-3Y (GluA2-3Y is in the -20 freezer in small tubes containing 0.9 to 2 mg, you have to adjust the amount of Tris-saline according to the amount written on the tube). Mix well and place the tube on ice.
     - Make aliquots of 20 µl and keep in the -20 freezer.     - Make aliquots of 20 µl and keep in the -20 freezer.
-    - Repeat the same procedure with the control peptide. +    - Repeat the same procedure with the control peptide (GluA2-3A). 
-  - Infusion solution 15pmol/ µl (to be prepared on same day of infusions)+  - **Infusion solution 15 pmol/ µl (to be prepared on same day of infusions)**
     - Thaw a tube of stock solution.     - Thaw a tube of stock solution.
     - Add 5 µl of stock solution to 995 µl of Tris-saline. Mix well.     - Add 5 µl of stock solution to 995 µl of Tris-saline. Mix well.
     - Discard the rest of stock solution remaining in the tube. Never refreeze a solution that has been thawed.     - Discard the rest of stock solution remaining in the tube. Never refreeze a solution that has been thawed.
     - Repeat the same with the control peptide (GluA2-3A)     - Repeat the same with the control peptide (GluA2-3A)
methods/sop/zipglua23y.1689357244.txt.gz · Last modified: 2023/07/15 15:20 (external edit)