Demeditec corticosterone rat mouse elisa

All mice were returned to their original cages after completing the MWM test (without any further stress) and were sacrificed 15 days later for blood and tissue sampling. Brain tissue was quickly removed and stored in a refrigerator for RNA-scope and western blotting.
Whole blood was collected via the eyeball. Serum was prepared by centrifuging blood samples for 5 min at 4000 rpm (4°C). Approximately 100 μl of serum was collected from each mouse and analyzed for corticosterone (CORT) levels with a quantitative ELISA (Demeditec Corticosterone rat/mouse ELISA, Demeditec Diagnostics, Germany). The MAP Mouse Cytokine/Chemokine Magnetic Bead Panel (Millipore, USA) was used to determine the levels of TNF-α and IL-1β.

Whole blood was collected via cardiac puncture and transferred into K₂EDTA microtubes for full and differential blood counts on the Cell-Dyne 3700CS hemocytometer (Abbott Diagnostics, USA). Plasma was collected from the whole blood samples and analyzed for basal corticosterone (CORT) concentrations by quantitative ELISA (Demeditec Corticosterone rat/mouse ELISA, Demeditec Diagnostics, Germany). Concentrations were calculated on a six-point standard curve with a logistic regression algorithm. The assay detection range was 6.1–2,250 ng/ml.
Spleens were collected in ice-cold complete RPMI 1640 (cRPMI) medium, consisting of 10% fetal bovine serum, 1% penicillin–streptomycin, and 1% gentamicin, for isolation of splenocytes.

Whole blood was collected by cardiac puncture and transferred to K₂EDTA microtubes. An aliquot was assigned for full blood and differential leukocyte counts on the CELL-DYN 3700CS haemocytometer (Abbott Diagnostics), while the remaining blood sample was used to collect plasma for assessment of corticosterone concentrations.
Corticosterone concentrations were determined by quantitative ELISA (Demeditec Corticosterone rat/mouse ELISA, Demeditec Diagnostics, Germany), as per manufacturer's instructions. The concentrations were calculated in Microsoft Excel using a 6-point standard curve with a logistic regression algorithm. The detection range of the kit was 6.1-2250 ng/ml. The kit has an intra-assay variation of 8.9% and interassay variation of 7.2%.
Mouse spleens were dissected under sterile conditions and collected into ice-cold complete RPMI 1640 medium (supplemented with 10% foetal bovine serum, 1% penicillin-streptomycin, and 1% gentamicin). Both LPS-treated dams and LPS-affected offspring displayed macroscopically visible larger spleen sizes in comparison to saline-treated dams or saline-affected offspring, respectively. Exact organ mass was however not determined due to the requirement for sterility in culturing splenocytes.