Streptozocin Injection.

v1
Procedures:
1. IMPORTANT: Inform Vivarium supervisor (Laurie Edwards at ltc8r@Virginia.EDU or Alice
Kenney at ask5g@virginia.edu, 434-924-5293) that you are going to perform STZ injection in
mice. Prepare a door label indicating the date, time and dose of the intended injection and hang it
on the door of the mouse room.
2. Remove the food from mice (DO NOT remove the waster bottle) and fast the mice overnight
starting around 5 pm (~16 hours).
3. Prepare Citrate buffer fresh in the morning by combining 20 ml of 0.1 M Sodium citrate with 30 ml
of 0.1 M Citrate acid to produce 0.1 M Citrate buffer. Adjust the pH to 4.0 by using 1 N NaOH.
Filter-sterilize the Citrate buffer and store it in a sterile conical tube on ice.
4. Weigh the mice and ear tag them.
5. Add Citrate buffer to Streptozocin (STZ) in a sterile conical tube in a biosafety hood right before
use to a final concentration of 20 mg/ml and keep on ice. For example, add 2.5 ml of Citrate buffer
to 50 mg of Streptozocin.
6. Inject STZ at 200 mg/kg, i.p. (0.1 ml/10 g).
7. Inject Citrate buffer at 0.1 ml/10 g for the control groups.
8. Feed the mice with 10% sucrose in place of water on the 1st day of injection.
9. Remove the sucrose and feed the mice with fresh water the next day.
10. Monitor body weight, blood glucose and ketone bodies after injection as needed.
11. Harvest tissue samples after euthanize the mice with CO2.

Reagent and solution preparation:

Streptozocin powder
Weigh Streptozocin (Hi-media) and store at -20C in aliquots (25-100 mg) in conical tubes that are
sealed well with cap and parafilm, and clearly labeled with the chemical name and weight.
0.1 M sodium citrate
Dissolve 1.47 g of sodium citrate tribasic dihydrate (Sigma C0909, MW 294.10) in 50 ml ddH2O
and store at room temperature.
0.1 M citric acid
Dissolve 1.05 g of citric acid monohydrate (Sigma C1909, MW 210.14) in 50 ml ddH2O and store
at room temperature.

Rat no.
01
02
03
04
05
06
Average
Diet (gm)
Water (ml)

Rat no.
01
02
03
04
05
06
Average
Diet (gm)
Water (ml)

Rat no.
01
02
03
04
05
06
Average
Diet (gm)
Water (ml)

Set. 1
Weight (gm)
Day 1 Day 2 Day 3
158
158
159
172
173
173
185
186
186
182
183
183
191
192
192
193
193
193
180.17 180.83
181
124
124
126
253
255
255

Set. 3
Weight (gm)
Day 1 Day 2 Day 3
184
184
184
199
199
199
209
209
210
210
210
210
197
198
198
191
191
191
198.33 198.5 198.67
128
128
132
256
254
258
Set. 5
Weight (gm)
Day 1 Day 2 Day 3
174
174
175
196
196
197
168
169
169
168
169
170
174
175
176
177
178
179
176.17 176.83 177.67
119
120
120
247
252
251

Rat no.
01
02
03
04
05
06
Average
Diet (gm)
Water (ml)

Rat no.
01
02
03
04
05
06
Average
Diet (gm)
Water (ml)

Rat no.
01
02
03
04
05
06
Average
Diet (gm)
Water (ml)

Set. 2
Weight (gm)
Day 1
Day 2
Day 3
191
191
191
168
169
169
198
198
199
203
203
203
210
210
211
189
189
189
193.17 193.33
193.67
127
130
128
250
252
256

Set. 4
Weight (gm)
Day 1
Day 2
Day 3
164
164
164
173
173
173
170
169
170
162
162
161
171
171
171
160
160
160
166.67
166.5
166.5
117
117
119
242
245
248
Set. 6
Weight (gm)
Day 1
Day 2
Day 3
175
176
176
169
169
169
170
170
171
157
158
157
181
182
182
170
171
171
170.33
171
171
123
127
125
254
257
254