The standard protocol is bedtime injection on an empty stomach to align with the bodys natural overnight growth hormone pulse window
If you do not track how much you have drawn from the vial, you have no way to know how much remains
(7)Fmoc-Ala-Asp(otBu)-Asp(otBu)-Ala-Gly-Leu-Val-WangResin (7) Preparation of Fmoc-Ala-Asp(otBu)-Asp(otBu)-Ala-Gly-Leu-Val-WangResin (6)Fmoc-Asp(otBu)-Asp(otBu)-Ala-Gly-Leu-Val-WangResin(20DMF)5203DMF3DMF2Fmoc-Ala-OH0.7gDIEA0.4mlTBTU0.56gHOBT0.2gDMF 10ml1.53DMF3DMF2Fmoc-Ala-Asp(otBu)-Asp(otBu)-Ala-Gly-Leu-Val-WangResin In the Fmoc-Asp(otBu)-Asp(otBu)-Ala-Gly-Leu-Val-WangResin in step (6), add a decapping reagent (20% piperidine in DMF solution), react at room temperature for 5 minutes, and drain, Re-add the capping agent and react at room temperature for 20 minutes, wash with isopropanol for 3 times, DMF for 3 times, re-distilled DMF for 2 times, drain, add Fmoc-Ala-OH0.7g, DIEA0.4ml, TBTU0.56g , HOBT0.2g, DMF 10ml normal temperature reaction 1.5 hours, drained with a vacuum pump, washed 3 times with isopropanol, washed 3 times with DMF, washed 2 times with DMF evaporated, and dried to obtain Fmoc-Ala-Asp(otBu )-Asp(otBu)-Ala-Gly-Leu-Val-WangResin
The ELISA kits used were bought from Thermo Fisher, USA, their detailed information is as follows: IL-4 Mouse Uncoated ELISA Kit (#88-7013-88), IL-5 Mouse Uncoated ELISA Kit (#88-7054-88), IL-13 Mouse Uncoated ELISA Kit (#88-7439-88), IL-17A (homodimer) Mouse Uncoated ELISA Kit (#88-7371-88 A), IL-23 Mouse Uncoated ELISA Kit (#88-7230-88), IL-6 Mouse Uncoated ELISA Kit (#88-7066-88), IL-33 Mouse Uncoated ELISA Kit (#88-7333-88) and TNF alpha Mouse Uncoated ELISA Kit (#88-7324-88)
