Mda elisa kit

CPG was measured using a URight blood glucometer (Uright, TD 4279) using the tail-prick technique to obtain tail vein blood. To collect fresh small medium of blood, an appropriate restraint device was used, and the withdrawal site was cleaned with alcohol. A finger prick lancet was used on the tail vein, and then capillary blood glucose was measured using a glucometer. After 12 d of treatment, the rat was euthanized before blood collection using cardiac puncture. The blood samples were then centrifuged at 4500 rpm for 10 min, and the serum samples were stored at 80°C until further analysis. Serum urea, creatinine, sodium (Na⁺), potassium (K⁺), calcium (Ca), magnesium (Mg), albumin, total protein, alanine transaminase (ALT), aspartate transaminase (AST), and alkaline phosphatase (ALP) were analyzed using an auto analyzer (Abbott ARCHITECT analyzer). Serum MDA was analyzed using the MDA ELISA Kit from Elabscience, USA, which uses the Sandwich-ELISA principle. All procedures were performed according to the manufacturer’s instructions.

Malondialdehyde (MDA) level was quantified from the rats’ serum strictly base on the simple principle of competitive-ELISA using MDA ELISA kit (E-EL-006, Elabscience). MDA level was assayed by monitoring the competitive reaction of the fixed amount of MDA on the pre-coated microtitter plate surface with the MDA in the serum samples. Standard working solution was set up for each sample well and 50 µL of the sample were added in each well separately. Immediately, biotinylated detection Ab working solution was added to each well sealed and incubated for 45 min at 37 °C. The solutions were aspirated and 350 µL of buffer was added to each well and soaked for 2 min then aspirated again 3 times before the addition of 100 µL of HRP conjugate working solution to each well and incubated for 30 min at 37 °C. Further, the incubated solution was decanted and washed with buffer solution 5 times before the addition of 90 µL of substrate reagent to each well and incubated for 15 min at 37 °C. Finally, 50 µL of stop solution was added to each well and the absorbance was measured using a micro-plate reader at a wavelength of 450 nm. The results were expressed as ng/mL.