rGHRH(1-29)NH2

Cell experiment:

To isolate GRF metabolites in the liver, rGRF(1-29)NH2 (10 mg) is first preincubated in 232 mL of Krebs' buffer (5 min, 37°C) to help GRF solubilization and then incubated with a liver homogenate in a shaking bath at 37°C. The homogenate is prepared as in the degradation assays with 580 mg of liver (10 mg/mL). The reaction is stopped after 30 min by adding 174 mL of cold 50 mM phosphate solution (pH 0.8) and centrifugation (48,000×g, 20 min, 4°C). The supernatant is filtered twice and its pH is adjusted (3.0) with 6 N NaOH before chromatography. The GRF metabolites and residual rGRF(1-29)NH2 are isolated[1].

References:

[1]. Boulanger L, et al. Catabolism of rat growth hormone-releasing factor(1-29) amide in rat serum and liver. Peptides. 1992 Jul-Aug;13(4):681-9.