All chemicals were obtained from Sigma unless otherwise stated. Antibodies to CREB (#9104), phospho S133 CREB (#9196), VASP (#3132), CRTC3 (#2720) and Lamin A/C (#4777) were from Cell Signaling Technologies. Antibodies to GAPDH (#MAB374) and CRTC2 (#ST1099) were from Merck-Millipore. Anti-BrdU antibody (B2531) was from Sigma Aldrich. Silencer® Select siRNA targeting CRCT2 and CRTC3 (#4390791 and #4390771 respectively) and negative control (#4390843) were purchased from Thermo Fisher. ON-TARGETplus SMARTpool CREB siRNA (#L-092995-02-0005) and ON-TARGETplus Non-targeting Control Pool (#D-001810-10-05) were purchased from Dharmacon. UO126, LY294002, SB 203580 were from Merck-Calbiochem. Bisindolylmaleimide I (BIS1, GF 109203×) was purchased from Tocris Bioscience. MK-2206 was from Selleckchem. HG-9-91-01 was a gift from Dr Kris Clark (University of Dundee)37 (link). Forskolin was purchased from Sigma Aldrich. BAY60-658350 (link) and Cicaprost were purchased from Caymen Chemical. PDGF-BB was purchased from R&D systems. 6-BNZ-cAMP-AM35 (link), a cell permeable selective PKA agonist, and 8-pCPT-2′-O-Me-cAMP-AM, a cell permeable selective EPAC super-agonist36 (link), were purchased from Biolog.
8 pcpt 2 o me camp am
Manufactured by Biolog
8-pCPT-2′-O-Me-cAMP-AM is a cell-permeant, light-activated cAMP analog. It acts as a selective activator of cAMP-dependent protein kinase (PKA).
Automatically generated - may contain errors
Lab products found in correlation
Silencer select sirna targeting, by Thermo Fisher Scientific (1 mentions)
Forskolin, by Merck Group (1 mentions)
Anti brdu antibody b2531, by Merck Group (1 mentions)
Lamin a c, by Cell Signaling Technology (1 mentions)
Pdgf bb, by R&D Systems (1 mentions)
On targetplus non targeting control pool, by Horizon Discovery (1 mentions)
Mk 2206, by Selleck Chemicals (1 mentions)
Sb203580, by Merck Group (1 mentions)
Ly294002, by Merck Group (1 mentions)
Uo126, by Merck Group (1 mentions)
Hoechst 33342 dye, by Thermo Fisher Scientific (1 mentions)
6 bnz camp am, by Biolog (1 mentions)
Axio observer, by Zeiss (1 mentions)
2 protocols using 8 pcpt 2 o me camp am
1
Signaling Pathway Analysis of CRTC Proteins
2
STIM1 Regulation of NFAT Translocation
Check if the same lab product or an alternative is used in the 5 most similar protocols
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 ).
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