Estradiol elisa kit

Whole blood samples were collected at the beginning of ovariectomy or sham surgery, at the beginning of ET-1 stroke induction surgery, and at the start of MRI sessions. Following clean venipuncture, we collected 300 to 500 μL of whole blood from the lateral tail vein immediately after induction with anesthesia. Samples were kept at 4°C for approximately 45 min to facilitate coagulation. Serum was isolated after centrifugation (21K rpm, 15 min, 4 C), then frozen at -20°C as individual aliquots for each rat and at each time point (serum was not pooled). Each sample of serum was divided into three 25 μL aliquot and assayed for Estradiol (Calbiotech Estradiol Elisa Kit, #ES180S-100) as per manufacturer’s instructions. The estradiol reaction was measured at 450 nm absorbance and relative fluorescence fitted to the reference standard curve.

Tartrate-resistant acid phosphatase (TRAP) activity (a marker of bone resorption) and serum estradiol (E2) levels were measured using a TRAP enzyme-linked immunoassay (ELISA) kit (USCN Life Science, Wuhan, China) and an estradiol ELISA kit (Calbiotech, San Diego, CA, USA), respectively. Bone alkaline phosphatase (BALP) levels were measured using a BALP ELISA kit (Elabscience, Wuhan, China).