6-Bnz-cAMP-AM, Me-cAMP-AM (8-pCPT-2′-O-Me-cAMP-AM), and ESI-09 were purchased from Biolog (Germany). Ryanodine, SKF-96365, and Xestospongin C were purchased from MilliporeSigma (Burlington, MA). Bt3-Ins(1,3,5)P3/AM was purchased from SiChem. ATP, 4-CMC (4-Chloro-m-cresol), H-89, and thapsigargin were from Sigma-Aldrich (St. Louis, MO).
6 bnz camp am
Manufactured by Biolog
Sourced in Germany
6-Bnz-cAMP-AM is a cell-permeant analogue of the second messenger cyclic AMP (cAMP). It can be used to study the effects of increased intracellular cAMP levels in a variety of cell types.
Automatically generated - may contain errors
Lab products found in correlation
Esi 09, by Biolog (1 mentions)
Adenosine triphosphate (atp), by Merck Group (1 mentions)
Thapsigargin, by Merck Group (1 mentions)
8 pcpt 2 o me camp am, by Biolog (1 mentions)
Hoechst 33342 dye, by Thermo Fisher Scientific (1 mentions)
Axio observer, by Zeiss (1 mentions)
Dibutyryl camp, by Merck Group (1 mentions)
Forskolin, by Merck Group (1 mentions)
3 protocols using 6 bnz camp am
1
Modulation of Intracellular Signaling Pathways
2
STIM1 Regulation of NFAT Translocation
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SH‐SY5Y STIM1−/− cells were transfected with STIM1‐mCherry, STIM1A‐mCherry, or noSTIM‐mCherry in combination with NFAT1‐GFP. 24 h post‐transfection, the GFP signal in the cytoplasm vs. in the nucleus was detected using the Zeiss AXIO observer. Immediately before the measurement, DNA was stained using 1 mg/ml Hoechst 33342 dye (Thermo Fisher Scientific). For inhibition of PDE8B, the specific blocker PF‐04957325 (1 µM, MCE) was added 2 h prior to the experiment and kept in all reagents during the measurement. To test the effect of cAMP analogs on NFAT translocation, cells were loaded 30 min prior to the measurement with 1 μM of 6‐Bnz‐cAMP‐AM (BioLog) or 8‐pCPT‐2'‐O‐Me‐cAMP‐AM (BioLog). The ratio of GFP signal in the cytoplasm vs. in the nucleus was determined using Fiji (Rueden et al, 2017 ).
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SH‐SY5Y STIM1−/− cells were transfected with STIM1‐mCherry, STIM1A‐mCherry, or noSTIM‐mCherry in combination with NFAT1‐GFP. 24 h post‐transfection, the GFP signal in the cytoplasm vs. in the nucleus was detected using the Zeiss AXIO observer. Immediately before the measurement, DNA was stained using 1 mg/ml Hoechst 33342 dye (Thermo Fisher Scientific). For inhibition of PDE8B, the specific blocker PF‐04957325 (1 µM, MCE) was added 2 h prior to the experiment and kept in all reagents during the measurement. To test the effect of cAMP analogs on NFAT translocation, cells were loaded 30 min prior to the measurement with 1 μM of 6‐Bnz‐cAMP‐AM (BioLog) or 8‐pCPT‐2'‐O‐Me‐cAMP‐AM (BioLog). The ratio of GFP signal in the cytoplasm vs. in the nucleus was determined using Fiji (Rueden et al, 2017 ).
3
EPAC1 agonist and PKA agonist protocol
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All chemicals were obtained from Sigma unless otherwise stated. Antibodies to EPAC1 (#4155) were from Cell Signalling Technologies. Antibodies to GAPDH (#MAB374) were from Merck-Millipore. Forskolin and dibutyryl-cAMP was purchased from Sigma Aldrich. BAY60-6583 was purchased from Cambridge Bioscience (U.K.). 6-BNZ-cAMP-AM [35] , a cell permeable selective PKA agonist, and 8-pCPT-2'-O-Me-cAMP-AM (8-CPT-AM), a cell permeable selective EPAC super-agonist [36] , were purchased from Biolog (Germany) and used at 20 µM in accordance with the manufacturers recommendations.
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