Ldh activity detection kit

We collected rat blood and left it at room temperature for 15 min. Then, we obtained the serum by centrifugation (3000 rpm, 15 min, 4°C). LDH activity detection kit (Sigma-Aldrich, St. Louis, MO, USA) and CK activity detection kit (Sigma-Aldrich, St. Louis, MO, USA) were used to detect the level of LDH and CK in serum. We used purified rat LDH antibody coating plate to make solid phase antibody. Then, we added unknown concentration of LDH test sample and known concentration of LDH standards to the monoclonal antibody. After incubating for 15 min, we added biotin-labeled anti-IgG antibody. The IgG antibody then binds to streptavidin-HRP to form an immune complex. Then, we added substrate TMB for color development. The color depth is positively correlated with the LDH in the sample. Finally, we used a microplate reader to measure the absorbance (OD value) at a wavelength of 450 nm and calculated the concentration of rat LDH in the sample through the standard curve. The same principle was also used to detect the level of CK.