These fractions were combined and concentrated with a Centricon Concentrator 10 (Millipore, Billerica, MA) to about 5 mL.
Peptide fractions were concentrated with a vacuum concentrator (Eppendorf), and kept in low-binding tubes at -20°C until further analysis.
The supernatant was concentrated with a gas blowing concentrator.
Total RNA was concentrated with a DNA100 SpeedVac Concentrator (Savant) and reverse transcribed using the High-Capacity cDNA reverse transcription (RT) kit (Applied Biosystems Inc., Foster City, CA).
The eluted fractions were filtered through a 0.22-µm filter and concentrated with a 5-kDa concentrator (Sigma-Aldrich, MO) by centrifugation at 4°C, washed and dialyzed against PBS.
The hybridoma supernatant was concentrated with a Vivacell 70 concentrator (Sartorius Biolab products) and purified with an NAb protein G antibody purification kit (Thermo Scientific) according to the manufacturer's instructions, and aliquots were stored at −20°C.
Peak fractions were pooled, concentrated with a stirred cell concentrator (EMD Millipore, Bilerica, MA) and dialyzed exhaustively against HEPES, pH 7.4, 150 mM NaCl.
The hybridization mix was concentrated with a Speed Vac Concentrator System (Eppendorf, Basel, Switzerland), resolved in 3× SSC, 0.3% SDS, denatured for 3 min at 95ºC, and incubated for 30 min at 37ºC before hybridization.
Dynabeads M-280 Streptavidin (Dynal) were concentrated with a magnetic particle concentrator (MPC) (Dynal) and washed twice in Buffer T (10 mM Tris (pH 7.5), 1 mM EDTA, 1 M NaCl).
The conditioned medium was collected, concentrated with a Centricon-10 (UFC910024, Billerica, MA), and protein concentrations were determined with the BCA assay (#23250, Pierce, Rockford, IL).
Similarly, co-crystallization with cAMP was accomplished by adding cAMP to a final concentration of 5 mM to the protein sample, which was concentrated with a 10 kDa cutoff Amicon Ultra (Millipore) to 17 mg/ml.
