Sections of the middle part of uterine horn collected from pigs were opened longitudinally on the mesometrial surface. Uteri were washed three times in sterile PBS then carefully cut into small pieces (400 mg weight) and then washed three times in medium M199 (Sigma, USA). Individual uterine slices were placed in culture vials containing 2 ml Medium 199 supplemented with 0.1% BSA (Sigma), 20 μg nystatin (Sigma) and 20 μg gentamicin (Krka, Novo Mesto, Slovenia) and then preincubated in vitro under atmosphere of 95% O2 and 5% CO2 at 37°C for 18 h. After preincubation, the culture medium was replaced with fresh medium, and the explants were treated with vehicle (control) or P4 (10−5 M; Sigma), E2 (10−9 M; Sigma), OT (10−7 M; Sigma), AA (10−5 M; Sigma), FSK (10 μg/mL; Sigma) and cpt-cAMP analog (200 μM; Sigma) and incubated for an additional 3 or 24 h. All treatments were performed in triplicates. Furthermore, uterine tissue explants were snap-frozen in liquid nitrogen (for RNA and protein extraction) and stored at −80°C until further use.
Cpt camp analog
Manufactured by Merck Group
Sourced in Slovenia
The Cpt-cAMP analog is a laboratory reagent used in biochemical research. It is a cyclic adenosine monophosphate (cAMP) analog that can be used as a tool to study cAMP-dependent signaling pathways. The core function of this product is to serve as a chemical compound for in vitro experimental applications.
Automatically generated - may contain errors
2 protocols using cpt camp analog
1
Uterine Tissue Explant Culture
2
Uterine Explant Culture and Treatment
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Forward: 5′-GACCTCCACTACATGGTCTA-3′
Reverse: 5′-AAGATGGTGATGGCCTTTC-3′
Uterine explants culture Sections of the middle part of uterine horn collected from pigs were opened longitudinally on the mesometrial surface. Uteri were washed three times in sterile PBS then carefully cut into small slices (400 mg weight) and then washed three times in medium M199 (Sigma, USA). Individual uterine slices were placed in culture vials containing 2 ml Medium 199 supplemented with 0.1 % BSA (Sigma), 20 µg nystatin (Sigma) and 20 µg gentamicin (Krka, Novo Mesto, Slovenia) and then incubated in a shaking water bath at 37 °C in a humidified atmosphere of 95 % O 2 and 5 % CO 2 for 18 h (Franczak et al. 2006) . After preincubation, the culture medium was replaced with fresh medium, and the explants were treated with vehicle (control) or P 4 (10 -5 M; Sigma), E 2 (10 -9 M; Sigma), OT (10 -7 M; Sigma), AA (10 -5 M; Sigma), FSK (10 µg/ml; Sigma) and cpt-cAMP analog (200 µM; Sigma) and incubated for an additional 3 or 24 h. Concentrations for the treatments were previously determined (Yang et al. 2003 , Franczak et al. 2006) . All treatments were performed in triplicates in five independent experiments. Furthermore, uterine tissue explants were snap-frozen in liquid nitrogen (for RNA and protein extraction) and stored at -80 °C until further use.
Reverse: 5′-AAGATGGTGATGGCCTTTC-3′
Uterine explants culture Sections of the middle part of uterine horn collected from pigs were opened longitudinally on the mesometrial surface. Uteri were washed three times in sterile PBS then carefully cut into small slices (400 mg weight) and then washed three times in medium M199 (Sigma, USA). Individual uterine slices were placed in culture vials containing 2 ml Medium 199 supplemented with 0.1 % BSA (Sigma), 20 µg nystatin (Sigma) and 20 µg gentamicin (Krka, Novo Mesto, Slovenia) and then incubated in a shaking water bath at 37 °C in a humidified atmosphere of 95 % O 2 and 5 % CO 2 for 18 h (Franczak et al. 2006) . After preincubation, the culture medium was replaced with fresh medium, and the explants were treated with vehicle (control) or P 4 (10 -5 M; Sigma), E 2 (10 -9 M; Sigma), OT (10 -7 M; Sigma), AA (10 -5 M; Sigma), FSK (10 µg/ml; Sigma) and cpt-cAMP analog (200 µM; Sigma) and incubated for an additional 3 or 24 h. Concentrations for the treatments were previously determined (Yang et al. 2003 , Franczak et al. 2006) . All treatments were performed in triplicates in five independent experiments. Furthermore, uterine tissue explants were snap-frozen in liquid nitrogen (for RNA and protein extraction) and stored at -80 °C until further use.
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